Organization, sequence and expression of the HLA

Immunobiology
E . D . A L B E R T , M ü n c h e n • H . D E I C H E R , H a n n o v e r • A . D E W E C K , B e r n • M . P. D l E R l C H ,
I n n s b r u c k • M . F E L D M A N N , L o n d o n • K . H A V E M A N N , M a r b u r g • S. H . E . KAUFMANN,
F r e i b u r g • H . K I R C H N E R , Heidelberg • E . K L E I N , Stockholm • W . KÖHLER, Jena • K . R E S C H ,
H a n n o v e r • G . RIETHMÜLLER, M ü n c h e n • M . RÖLLINGHOFF, Erlangen • K . O . R O T H E R ,
H e i d e l b e r g • D . S C H E N D E L , M ü n c h e n • V . SCHIRRMACHER, Heidelberg • C . SORG, M ü n s t e r •
R . T l M P L , M ü n c h e n • R . VAN F U R T H , L e i d e n • H . W A G N E R , U l m • G . W l C K , I n n s b r u c k •
R. ZlNKERNAGEL, Zürich
Editor-in-Chief
D . G E M S A , Marburg
Editorial Advisory Board
R . A V E R D U N K , B e r l i n • J . F . B A C H , Paris • H . B A L N E R , R i j s w i j k • R . B E N N E R , Rotterdam •
D . B l T T E R - S U E R M A N N , M a i n z • H . v. B O E H M E R , Basel • G . B O N N A R D , B e r n • D . G .
B R A U N , Basel • V . B R A U N , T ü b i n g e n • J . BROSTOFF, L o n d o n • A . C O U T I N H O , Paris •
T . D I A M A N T S T E I N , B e r l i n • W . D R Ö G E , Heidelberg • P . D U K O R , Basel • P. E R B , Basel •
H . - D . F L A D , Borstel • O . G Ö T Z E , G ö t t i n g e n • E . G Ü N T H E R , F r e i b u r g • U . H A D D I N G , M a i n z •
H . H A H N , Berlin • K . H Ä L A , I n n s b r u c k • G . J . HÄMMERLING, Heidelberg • K . U . H A R T MANN, M a r b u r g • H . ZUR H A U S E N , Heidelberg • M . HESS, B e r n • J . K A L D E N , Erlangen •
B . K I N D R E D , T ü b i n g e n • T . J . K I N D T , N e w Y o r k • U . KOSZINOWSKI, T ü b i n g e n • E . K O W NATZKI, F r e i b u r g • P . KRAMMER, Heidelberg • W . L E I B O L D , H a n n o v e r • K . L E N N E R T , K i e l •
F . L I L L Y , N e w Y o r k • J . L I N D E M A N N , Z ü r i c h • E . M A C H E R , M ü n s t e r • H . M E T Z G E R , Bethesda •
V . TER M E U L E N , W ü r z b u r g • H . J . MÜLLER-EBERHARD, L a Jolla • W . MÜLLER-RUCHHOLTZ,
K i e l • H . H . P E T E R , F r e i b u r g • H . PETERS, G ö t t i n g e n • E . P I C K , T e l A v i v • O . PROKOP, Berlin •
M . Q U A S T E L , Beer Sheva • J . P. R E V I L L A R D , L y o n • E . P. R l E B E R , M ü n c h e n • E . RÜDE, M a i n z
• E . SCHÖPF, Freiburg • H . G . SCHWICK, M a r b u r g • K . SETHI, Heidelberg • G . SUNSHINE,
L o n d o n • N . T A L A L , San Francisco • G . T I L L , A n n A r b o r • G . U H L E N B R U C H K ö l n •
M . W A G N E R , Jena • H . W E K E R L E , W ü r z b u r g • P . W E R N E T , T ü b i n g e n
Volume 170
Gustav Fischer Verlag • Stuttgart • New York • 1985
UniversitätsBibliothek
München
I S S N Immuno biology • Zeitschrift für Immunitätsforschung • 0171-2985
© Gustav Fischer Verlag • Stuttgart • New York • 1985
Alle Rechte vorbehalten
Printed by Druckerei Ungeheuer + Ulmer K G G m b H + C o , Ludwigsburg
Printed in Germany
Immunobiol., vol. 170, pp. 367-380 (1985)
Original Papers
Institute of Immunology, University of Munich, Federal Republic of Germany
Organization, Sequence and Expression of the HLA-B27
Gene: A Molecular Approach to Analyze HLA and Disease
Associations
E L I S A B E T H H . WEISS, W . K U O N , C . D Ö R N E R , M . L A N G ,
and G. R I E T H M Ü L L E R
Received November 1,1985 • Accepted November 14,1985
Abstract
A m o n g the numerous autoimmune diseases associated with various H L A alleles, the one
w i t h the highest relative risk so far reported has been ankylosing spondylitis w i t h H L A - B 2 7 .
To examine this relationship more directly, we have cloned the gene encoding the H L A - B 2 7
antigen and determined its complete D N A sequence. Comparison of the H L A - B 2 7 sequence
w i t h that of the allelic H L A - B 7 shows a high level of homology. Mutations are distributed
evenly between exons and introns. Exon 1 and intron 1 are the most divergent ones, and the
degree of divergence distinctly declines towards the 3' end. The H L A - B 2 7 gene when
transfected into murine L cells is expressed on the cell surface and reacts with a panel of
monoclonal antibodies directed against monomorphic and polymorphic determinants associated with H L A - B 2 7 antigen.
The isolation of this gene allows for the first time a search for structural features which make
the H L A - B 2 7 antigen a high risk genetic factor for a group of rheumatoid disorders, in
particular ankylosing spondylitis.
Introduction
Among the numerous H L A markers associated with various human
diseases, the H L A - B 2 7 antigen is a clear exception. This allelic glycoprotein, present in the cell membrane of all nucleated cells, represents an
unusually high-risk factor for its carriers, conveying susceptibility for a
group of joint diseases of which ankylosing spondylitis (AS) is the most
prominent. Pooling all data on Caucasians, one arrives at a relative risk of
more than 70.0 for individuals carrying the H L A - B 2 7 antigen (1). Recent
Abbreviations: AS = ankylosing spondylitis; bp = base pair; kb = kilo bases; mAb =
monoclonal antibody; T = cytotoxic T cell; tk = thymidine kinase.
c
368
• ELISABETH H . WEISS, W . K U O N , C . DÖRNER, M . L A N G , and G . RIETHMÜLLER
studies have been directed towards elucidating the biochemical structure of
the H L A - B 2 7 antigen (2, 3). So far, no differences between the H L A - B 2 7
antigens present in healthy and AS patients have been found. Even the
subdivision of H L A - B 2 7 into several antigen subtypes (4) did not indicate
any structural differences between the H L A - B 2 7 molecules from healthy
and affected individuals.
In order to investigate the mechanisms of H L A involvement in disease, in
general, and the nature of the H L A - B 2 7 and AS association in particular,
we isolated the gene encoding the H L A - B 2 7 antigen. In this paper we
describe the expression of the H L A - B 2 7 gene in murine L-cells and present
the complete D N A sequence of the gene encoding the H L A - B 2 7 polypeptide together with its deduced complete amino acid sequence. The genomic
organization of the H L A - B 2 7 gene is compared with a cross-reactive allele,
the homologue H L A - B 7 gene, which exhibits no significant disease association.
Materials and Methods
Isolation of the HLA-B27
gene
A genomic library was constructed w i t h the vector p T C F (5, 6 ) from peripheral white blood
cells of a healthy individual with the H L A type: H L A - A 2 / 2 , -B5/27, -Cw2/3. The cosmid
library was screened w i t h an H L A - B locus specific c D N A probe derived from the 3'untranslated region of the H L A - B 8 gene (7).
DNA
sequence analysis
D N A sequence analysis of the three B g l l l fragments containing the entire H L A - B 2 7 gene
subcloned in the vector p U C l 3 was carried out mostly by the dideoxy sequencing method (8)
and partly, to verify ambiguous nucleotides, by the chemical sequencing procedure (9). The
sequencing strategy is shown in Figure 1.
Transfection of cosmid clone CD2.6 into murine Ltk~ cells
Murine thymidine kinase negative (tk~) cells, L D l ( H - 2 ) were transformed with cosmid
D N A and the herpes tk gene contained in the vector p O P F (5), using the calcium phosphatemediated D N A transfer technique (10). Populations of transformed cells were selected using
hypoxanthine-aminopterin-thymidine ( H A T ) selection, and stable L t k transformed clones
were established. The human lymphoblastoid cell line L G - 2 ( H L A - A 2 , 2 ; B27/27; Cwl/1) was
used as a positive control for H L A - B 2 7 expression.
k
+
Monoclonal
antibodies (mAb)
The origin and characteristics of the mAbs used are listed in Table 1.
Immunofluorescence
staining and
cytofluorography
Cell surface antigen expression by L cells was studied by EPICS analysis (EPICS V , Coulter
Electronics). Monolayer cultures of L cells were harvested by 1 m M PBS/EDTA and washed
three times in PBS, 0.1 % N a Azide, 10% FCS. Approximately 10 -10 cells were incubated
w i t h 50 u.1 of culture supernatant or ascitic fluid (1:10 ) of monoclonal antibodies. After 1-h
5
5
6
Organization and Expression of the H L A - B 2 7 Gene • 369
CD 2.6 J _
HLA-B27
exon
1
6
2
T
7
8/3'ut
t T
T
T
5 0 0 bps
T
T ?
Fig. 1. Restriction map of the cosmid clone CD2.6, fine restriction map of the H L A - B 2 7 locus
and sequence strategy.
T o p line: Restriction map for the cosmid clone CD2.6 encoding the H L A - B 2 7 antigen. The
following restriction sites are shown: K = K p n l ; S = Sali; X = X h o l . Middle line: The fine
restriction map of the H L A - B 2 7 locus, t = B g l l l ; T = K p n l ; ? = PstI; 7 = P v u I I ; T =
Sacl. Above the restriction map is shown the position of the exons and the 3' untranslated
region (3' ut).
a) sequences determined by the dideoxy sequencing method (8).
b) sequences obtained by the chemical sequencing procedure (9).
incubation, cells were washed and stained with 50 jil of fluorescein-conjugated goat antimouse I g G + I g M (Jackson Immunoresearch Laboratories, Inc.). Fluorescence intensity was
measured by flow microfluorimetry by analyzing 1-2 x 10 stained cells. Fluorescence data for
the expression of H L A - B 2 7 antigen on the transfected L cells are expressed as fluorescence
intensity relative to the L cell of the native H - 2 phenotype.
4
k
Table 1. Characteristics of the monoclonal antibodies used
Monoclonal""
antibodies
Ig class
Specificity
References
W6/32
11-4.1
MPC11
KTl
MA2.1
ME1
BB7.1
B27M2
IgG2a
IgG2a
IgG2b
IgM
IgG,
IgG,
IgG,
IgM
anti H L A - A , B , C
anti H - 2
undefined
anti H L A - A
anti H L A - A 2 , B17
anti H L A - B 7 , B27, B22
anti H L A - B 7
anti H L A - B 2 7 , Bw47
20
21
22
23
24
25
26
2
k
* Monoclonal antibodies ref. 20-26 were derived from culture supernatants and were gifts
from D r . J . P. Johnson. Antibody B27M2 (Ascites, 1:10 ) was a gift from D r . F. C. Grumet.
5
370
• ELISABETH H . WEISS, W . K U O N , C . DÖRNER, M . L A N G , and G . RIETHMÜLLER
Results
Genomic organization of the HLA-B27
gene
The entire H L A - B 2 7 gene is contained in the cosmid clone C D 2.6. The
restriction map of the cosmid clone and the gene, itself contained in three
B g l l l fragments, is shown in Figure 1. No other class I gene was located
within a distance of 6 kb to the left side and of 30 kb to the right side of the
H L A - B 2 7 locus. Since this cosmid clone contains only one H L A class I
gene, it was used directly for the gene transfer experiments.
Expression of the human HLA-B27 antigen in murine cells transformed
with the cloned HLA-B27 gene (CD2.6)
We have introduced cosmid C D 2.6 (see Fig. 1) together with the herpes
simplex virus thymidinekinase (tk) gene into mouse L tk~ (of H - 2
haplotype) cells using calcium phosphate D N A transfer. We tested the tk
k
MPC11
s
LD1
IAJ6/32
3
z
MPC11
0)
2.6
O
B
IAJ6/32
11-4.1
i
Relative
Fluorescence
Fig. 2A and B. EPICS-Analysis: Reactivity of the L D l cell line (Fig. 2A), and the transfection
clone 2.6 (Fig. 2B) w i t h different anti-H-2 and a n t i - H L A monoclonal antibodies. Fig. 2A
represents the fluorescence profiles of the L D l cell line ( H - 2 ) with the mouse plasmocytoma
antibody M P C - 1 1 and monoclonal antibodies W6/32 ( H L A - A , B, C) and 11-4.1 ( H - 2 ) . Fig.
2B shows the reactivity pattern of the transfection clone 2.6 w i t h the same Moabs M P C - 1 1 ,
W6/32 and 11-4.1. The fluorescence profiles were obtained by analysing 2 x 10 cells.
Fluorescence intensity was assessed by flow microfluorimetry by using integral amplification.
k
k
4
Organization and Expression of the H L A - B 2 7 Gene • 371
Table 2 \ Binding of anti-H-2 and a n t i - H L A antibodies to the mouse L cell line L D l , the
mouse L cell transfectant 2.6 and the human B cell line L G - 2
:
Cell
LDl
LDl
LDl
LDl
2.6
2.6
2.6
2.6
2.6
2.6
2.6
2.6
2.6
LG-2
LG-2
LG-2
LG-2
LG-2
LG-2
LG-2
LG-2
LG-2
Specificity
% pos.
cells
Fluorescence
intensity
(median)
11-4.1
MPCll
W6/32
H-2
undefined
HLA-A,B,C
1.7
95.1
3.4
2
4.3
140.0
5.8
4.3
11-4.1
MPC11
W6/32
KT1
MA2.1
ME1
BB7.1
B27.M2
H-2
undefined
HLA-A,B,C
HLA-A
H L A - A 2 , B17
H L A - B 7 , B27, B22
HLA-B7
H L A - B 2 7 , Bw47
1.4
98.0
2.3
96.0
1.4
1.9
95.0
2.3
9.3
4.8
150.0
5.1
50.0
4.6
4.6
40.4
4.8
10.4
HLA-A,JB,C
H-2
undefined
HLA-A
H L A - A 2 , B17
H L A - B 7 , B27, B22
HLA-B7
H L A - B 2 7 , Bw47
3.0
88.0
2.0
4.0
60.0
68.0
58.8
18.0
49.5
9.1
173.0
6.3
8.2
41.8
47.7
138.1
11.3
90.0
Antibody
_
k
k
_
W6/32
11-4.1
MPC11
KT1
MA2.1
ME1
BB7.1
B27.M2
k
" The results are representative of several different experiments
::
positive clones for expression of H L A class I cell surface antigens, using
monoclonal antibody binding in immunofluorescence assays.
The results of the binding assays are shown in Figure 2 and Table 2. All
clones derived from the CD2.6 transformation bind anti-HLA class I
monomorphic antibody (W6/32) at a level comparable with the endogenous
H-2 antigen expression. The level of H L A antigen detected was 30-50 % of
that of the H - 2 antigen.
These assays demonstrate that L cells transformed with cosmid CD2.6
expressed a cell-surface polypeptide which is recognized by W6/32, while
the untransformed cells were completely negative. To characterize the
HLA-antigen encoded by the clone CD2.6 we used several monoclonal
reagents directed against H L A antigens of the donor from whom the
cosmid library was constructed (see Table 2). The H L A - A 2 allele was
excluded, since the monomorphic anti-HLA-A monoclonal antibody, K T l ,
and monoclonal antibody MA2.1, which reacts with H L A - A 2 , did not bind
to the transfectant clones (or gave signals comparable to the M P C l l
control). The cosmid clone CD2.6 did bind, however, two monoclonal
k
372
• ELISABETH H . WEISS, W . K U O N , C. D Ö R N E R , M . L A N G , and G . RIETHMÜLLER
antibodies directed against HLA-B27. The reagent M E l binds to the
transfectants at the same level as W6/32. This binding is not due to crossreactivity with H L A - B 7 , since binding of BB7.1 was negative. The monoclonal antibody B27.M2, which recognizes a subset of H L A - B 2 7 antigens,
also binds to the transfectant clones, although at a much lower level. This
reagent is strongly positive for a B cell line derived from the individual from
whom CD2.6 was isolated. The lower binding of this mAb to the transfected L cell may be due to the combination of xenogeneic ß microglobulin
with the H L A - B 2 7 polypeptide. The expression of transfected H L A - B 2 7
gene in the murine cells is increased twofold after incubation with lymphognes present in a supernatant derived from rat spleen cells stimulated
with Concanavalin A (data not shown). These data show that cell lines
transformed with cosmid CD2.6 express a cell-surface antigen with a
specificity which is indistinguishable from the H L A - B 2 7 antigen of the
positive control B cell line L G - 2 (HLA-A2,2; B27/27). We therefore
conclude that the clone CD2.6 codes for the H L A - B 2 7 antigen.
We tried to determine whether the presence of the H L A - B 2 7 molecule
on the murine cells could render these cells as targets for allogeneic
cytotoxic T cell ( T cell)-mediated killing by human T cells raised against
the H L A - B 2 7 antigen. Thus far, however, human anti-B27 T cells did not
lyse the transfectant L cell clones significantly (data not shown). It was
reported previously that murine L cells expressing H L A class I genes are
poor targets for human allogeneic T cells (11, 12), whereas other transfected murine cells expressing H L A class I antigens can be lysed by human
allogeneic T cells (12).
2
c
c
c
c
c
General structure of the HLA-B27
gene
The overall structure and dimensions of the H L A - B 2 7 gene are the same
as those of other reported H L A genes. The H L A - B 2 7 gene consists of eight
exons and seven introns. Table 3 shows that the majority of the exons are of
the same length as those of the H L A - B 7 (14) and H L A - A 2 (14, 15) genes.
The sequence of cloned H L A - B 2 7 c D N A derived from the L G 2 line, which
we reported recently, confirms these exon assignments (16). The approximate size of the introns is the same in all three genes, with only minor
variations in the exact lengths.
DNA sequence of the HLA-B27
gene
We determined the D N A sequence of the entire H L A - B 2 7 gene. The
sequencing strategy is shown in Figure 1. The complete D N A sequence
together with the deduced amino acid sequence of the HLA-B27 antigen is
shown in Figure 3.
The sequence of the H L A - B 2 7 gene is similar to that of other human
class I genes reported earlier (14-19). For the first time, determination of
the exact length of exon 1 could be made from the sequence of this H L A -
Organization and Expression of the H L A - B 2 7 Gene • 373
Table 3. Sequence comparison of class I genes
Region
Length (bp)
H L A - -B27 B 7
A2
b
5' region
exon 1
intron 1
exon 2
intron 2
exon 3
intron 3
exon 4
intron 4
exon 5
intron 5
exon 6
intron 6
exon 7
intron 7
8 + 3' ut
3' flanking
a
b
c
d
e
517
73
129
270
241
276
575
276
92
117
442
33
106
48
182
424
209
507
73
123
270
245
276
575
276
92
117
442
33
106
48
182
422
209
% divergence
B27/A2
A2/A3
3
C
526
73
129
270
240
276
599
276
97
117
436
33
142
48
169
405
160
B27/B7
8.1
10.9
9.3
5.2
5.4
6.1
1.0
0.4
1.1
0.8
2.2
0
1.9
0
2.7
1.8
1.9
10.6
4.1
10.9
3.0
3.3
4.7
7.7
4.0
3.1
2.6
5.0
3.0
2.1
2.1
4.7
0.5
5.5
10.2
15.0
12.4
10.7
9.5
8.3
15.8
10.1
20.6
14.5
14.5
12.2
16.9
12.5
23
nd
nd
e
H-2K /H-2K
b
d
nd
7.8
3.7
10.3
4.3
11.2
4.0
6.1
4.7
5.8
3.9
6.0
2.3
2.6
1.8
5.0
4.0
Insertions and deletions were counted as one change
The sequence of Biro et al. (14)
The sequence of Koller et al. (15)
The comparison of Weiss et al. (29)
Calculated for 165 bps of the H L A - A 2 gene
nd = not determined
B27 gene. All other H L A class I genes published to date contain two in
frame initiation codons (see Fig. 4) which would result in two possible
leader peptides of 21 and 24 amino acids. HLA-B27 contains only the first
A T G with an exon 1 of 49 base pairs. The gene contains the 3.5 kb Taq I
fragment, a previously described restriction fragment length polymorphism
which was assigned to the H L A - B 2 7 gene (27). The fragment was detected
by using an H L A class I c D N A probe. The Taq I fragment spans the 3' half
of the H L A - B 2 7 gene. The detection of this HLA-B27-specific gene band
becomes easier when a probe derived directly from this region or an H L A B locus specific probe is used.
Potential Alternative Splicing Signals in the HLA-B27 gene. Recently, an
alternative intron/exon organization affecting the second exon has been
proposed for the H - 2 K gene (28). In a c D N A clone, apparently derived
from a processed H - 2 K transcript, an alternative splice acceptor site in the
first intron, 50 nucleotides 5' to the usual acceptor site, is used. In addition,
an extra intron is created by the splicing out of a D N A segment of exon 2,
coding for the amino acids 6-38 of the classical H - 2 K antigen. Until now it
could not be shown whether this alternative splice site is used in vivo
yielding a polypeptide that is synthesized from this alternative m R N A
transcript.
d
d
d
GACCTCACTCTCTCGCATCAACTTCTCCCTCATCACTTTCCCTACACAACATCCAIGACCACÄCCTAAGCACTCACACGCAGGCAGTCCACTTCACCGACAGGCATTCCACCACCACiAGTGAACGCGAACCCGCTCCCCGCCACTCCCC
CTCTCTCCCTGGTTTCCACAGACACATCCTTGTCCCGGACTCAGCCAGACAGTCTCACAAACACGCTGGTGTAGCACA
ACACGGATCAGCACGAACGTCCAAGCCCCCGGCCGCGCTCTCAGCCTCTCAGCCTCCGACACCCTTCTCTGC
ATTGGCGACGCGCACAGTTCCGGATTCCCCACTCCCACGAGTTTCACTTCTTCTCCCAACCTATGTCGGCTCCTTCTTCCiCGATACTCCTGACCCGTCCCCATTTCCCACTCCCATTGGGTCTCGCCTCTCT
AGAGAACCCAATCACTC
TCGCCCCGCTCCCACTTCTAAAGTCCCCACCCACCCACCCCCACTCACAATCTCCTCACACGCCGAC
« e t Arg V a l T h r A l a P r o Arg T h r L e u L e u L e u L e u L e u T r p G l y A l a V a l A l a Leu Thr G l u
ATC CCC CTC ACC CCC CCC CCA ACC CTC CTC CTG CTG CTC TCC CCC GCA CTG CCC CTC ACC GAG
Thr Trp Ala C
ACC TCC GCT C GTGAGTCCCCGCTCAGCCAGCGAAATCCCCTCTCTCCCGACCAGCCACGCCACCCACGCCCCGGCGCAGCACCCGCGGAGCCCCCCCCGCACCACGGTCCCGCCGCTCTCACCCCCTCCTCCCCCCCAC
His
ly Ser
CC TCC
,$
0
1,53
588
72M
L,
S e r « e t Arg T y r Phe H i s T h r S e r T a l S e r A r g P r o C l y Arg C l y C l u P r o A r g Phe l i e T h r V a l C l y T y r V a l Asp Asp T h r L e u Phe V a l Arg Phe A l a S e r Asp A l a
CAC TCC ATC AGG TAT TTC CAC ACC TCC CTC TCC CCC CCC CCC CCC CCG CAC CCC CGC TTC ATC ACC GTG CCC TAC CTG CAC CAC ACC CTG TTC CTC AGG TTC CAC ACC CAC CCC
838
A l a S e r P r o Arg G l u G l u P r o Arg A l a P r o t r p l i e G l u C l n C l u C l y P r o G l u T y r T r p Asp Arg G l u T h r G i n l i e C y s L y s A l a L y s A l a C l n T h r Asp Arg G l u Asp Leu
CCG ACT CCG AGA CAC CAC CCC CCC CCG CCG TCC ATA CAC CAC GAG CGC CCG CAC TAT TCC CAC CCC GAG ACA CAC ATC TCC AAG CCC AAG GCA CAG ACT GAC CCA CAC GAC CTC
<J$2
Arg T h r Leu L e u Arg T y r T y r Asn C l u S e r C l u A l a C
CCG ACC CTC CTC CGC TAC TAC AAC CAC ACC CAC CCC C CTGACTCACCCCCCCCCCGCCGCACCTCACGACTCCCCATCCCCCACCTACGGCCCGCCTCCCCCCCÄGTCTCCGGCTCCGÄCATCCCCCCCCGAGCCCG
CCCCACCCGCCCACACCCTCCACCCCCCACACCCCACGCGCCTTTACCCCCTTTCATTTTCACTTCACGCCAAAATCCCCCCCGTTGGTCCCCCCGCCCCGCGGCTCGGGCCdCACCCGGCTGACCCCCGGCCCCCCTCCAC
l y S e r B i s T h r Leu C l n Asn « e t T y r C l y C y s Asp V a l G l y P r o Asp C l y Arg L e u Leu Arg C l y T y r H i s G i n Asp A l a T y r Asp G l y L y s Asp T y r l i e A l a Leu Asn G l u
GC TCT CAC ACC CTC CAG AAT ATC TAT CCC TCC CAC CTC CCC CCC CAC CGC CCC CTC CTC CCC CCG TAC CAC CAG GAC CCC TAC CAC GGC AAG CAT TAC ATC CCC CTC AAC CAG
03
PI
ieß9
^
1 3«3
2
v>
^
Asp Leu S e r S e r T r p T h r A l a A l a Asp T h r A l a A l a G i n H e T h r G i n Arg L y s T r p C l u A l a A l a Arg V a l A l a C l u C l n Leu Arg A l a T y r L e u G l u C l y C l u C y s Val C l u
GAC CTG ACC TCC TGG ACC CCC CCG CAC ACG CCG CCT CAG ATC ACC CAG CGC AAG TCC GAG GCC CCC CGT GTG GCG CAC CAG CTC AGA GCC TAC CTG GAG CCC CAG TCC CTG CAG
T r p L e u Arg Arg T y r L e u G l u Asn C l y L y s C l u T h r L e u C l n Arg A l a A
TGG CTC CCC AGA TAC CTC CAC AAC CGC AAG CAG ACC CTC CAC CGC GCC C GTACCAGGGCCAGTGCGCACCCTTCCCCATCTCCTATAGGTCCCCCCCCATCCCCTCCCACCACAACACCAGGAAAATCCCATC
ACCCCTACAATCTCCCCCTCCCTTGAATGCAGAATCGCATCACTTTTCCTG1GTTTCCTCTCACGGCCCCCTCTTCTCTCTAGGACAATTAAGGGATCACGTCTCTGAGGAAATGGACCGCAAGACACTCCCTACAATACTGATCACCSO
TCCCCTTTGACCCCTGCAGCAGCCTTGCCAACCCTGACTTTTCCTCTCACGCCTTGTTCTCTGCCTCACACTCAGTCTGTTTCGGGCTCTCATTCCACCACTTCTGACTCACTTTACCTCCACTCAGATCACCAGCAGAAGTCCCTCTTC
CCCCCTCACACACTCCAACTTTCCAATGAATACCAGATTATCCCACCTCCCTCCGTCCACGCTCCTCTCTCCGTTCTGTGCCCCTTCCCCACCCCACCTCTCCTCTCCATTCTCAGCCTGGTCACATCCGTCCTCCTACCCTCTCCCATC
AGAGATGCAAAGCGCCTCAATTTTCTCACTCTTCCCATCAG
s p P r o P r o L y s T h r B i s V a l T h r B i s B i s P r o H e S e r Asp H i s C l u A l a T h r L e u Arg C y s T r p A l a Leu C l y Phe T y r P r o
AC CCC CCA AAG ACA CAC GTC ACC CAC CAC CCC ATC TCT GAC CAT CAC CCC ACC CTG ACC TCC TCC CCC CTC CGC TTC TAC CCT
1«»S7
1598
Q
1898
^
21 bu
A l a C l u H e T h r L e u T h r T r p C l n Arg Asp C l y C l u Asp C l n T h r C l n Asp T h r C l u L e u V a l C l u T h r Arg P r o A l a C l y Asp Arg T h r Phe C l n L y s T r p A l a A l a V a l V a l
GCG CAC ATC ACA CTC ACC TGC CAC CCC GAT CCC CAC GAC CAA ACT CAC CAC ACT GAC CTT GTC CAG ACC ACA CCA GCA CCA CAT ACA ACC TTC CAG AAC TCC CCA CCT GTC CTC
22?8
T a l P r o S e r G l y C l u C l u C l n Arg T y r T h r C y s H i s V a l C l n B i s C l u G l y L e u P r o L y s P r o L e u T h r L e u Arg T r p G
GTG CCT TCT GCA CAA GAG CAC AGA TAC ACA TCC CAT CTA CAC CAT CAC CGG CTC CCC AAC CCC CTC ACC CTC ACA TGG G GTAAGGACCCGCATCAGCCCTCATATCTCTTCTCACCCAA
2U80
ACCACCCCTTCACCAGGTCACGGCCCCTCATCTTCCCTTCCTTTCCCAC
!*
Q
:
PS
2j
PI
^
ACC
l u P r o S e r S e r C l n S e r T h r V a l P r o l i e V a l G l y l i e V a l A l a C l y L e u I I a V a l Leu A l a V a l V a l V a l H e C l y
AG CCC TCT TCC CAG TCC ACC CTC CCC ATC GTG CCC ATT GTT OCT GGC CTG OCT CTC CTA CCA CTT GTC CTC ATC CCA
>
z
A l a V a l V a l A l a A l a T a l n e t C y s Arg Arg L y s S e r S e r G
GCT GTC GTC OCT CCT CTG ATC TGT ACC AGG AAG ACC TCA C
GTACGGAACGGGTGAGGCGTCCCCTCTCAGTTTTCTTGTCCCACTCGCGGTTTCAACCCCCACCTACAAGTGTTCCCTCCCTCATTACTCCCAACCA
GCATCCACACACCGGCTAACGCACCCTCGCACCCTGTGTCCCACCACTTACTCTTTTCTGCACCACATCTCACAATCAACCACCCATGTATCACCTTCGTGGTTCTCGTGTTCGGGTCCTCATTCCACCATTCATGACTCACCGGAACCTC
CCTGCTAAGCACAGACCTTAGGACCGCAGTTGCTCCAGCACCCACACTTCCTTTCCTCGTCTTTCCTGATCCTGCCTTGGGTCTGTAGTCATACTTCTGCAAATTCCTTTTGGGTCCAACACGACCAGGTTCCTCTAACATCTCATGCCCC
TCCTTCCTCCCACTCCCCTCACACGCAAATTTTCTTCCCACAC
ly
C l y Lys Cly Gly Ser Tyr Ser Gin Ala Ala C
CT
CCA
ys
TCTCCTGCGCCCTCTGACCACGTCCTCTTTTTCTTCTACTCCAC
Ser
AAA
Asp
G C A GCC
Ser
Ala
ACC
Cln
T A C TCT
Cly
Ser
CAC
Asp
C C T GCC
Val
Ser
T GTAAGTGATGCGGGTCGGACTGTGGAGGAGCTCACCCACCCCCTAATTCCTCCTGTCCCACG
Leu
Thr
Ala
CC ACC G A C A C T CCC C A G CCC TCT C A T CTC TCT CTC ACA CCT T C A AAA
3230
X
3535
Q
C C TCACAC
AACCCACCTCAATGTCTCTCCGTCCCTCTTACCATA ATGTCACCACGTCCAGAGACCAGCCCACCC
2
CCGTCTCCACTCTGACCCCTCTTCCCATGCTGACCTGTGTTTCCTCCCCACTCATCTTTCCTCTTCCACACACCTCCGCCTCGATGTCTCCATCTCTCTCTCAACTTTATCTCCACTCACCTGCAACTTCTTACTTCCCTACTCAAAATA
AGAATCTGAATATAAATTTCTTTTCTCAAATATTTCCTATCACAGCTTCATGGATTAATTAAATAAGTCAATTCCTCGAATTTCAGACACCAAATAAACACCTCACAACCTTCCAGAATCTGCATCTTCGCTCTGCTCACTCTGTTCCAC
p
38 35
CTCGCCTGTCCAGAAGGCTCTGCCCCGCCCAGTCTGCACCCCCCTCTGCCCATTTCGTGTTCACTCCATCATCGCCTTTATCTCCTTACTCCTCACCTCCCTCACCTTCACTCCTCCATTCTCCTTGTCCCTTCACTCCAAACGTTGTCC
ACCGCGACCTGTCACCACACAGCCTCACAC
p0
£j
G CTCACATTCTTCCCCTCTACACTCCCTCCCCTGGCACCTCT
GGCCCTCCCTCCCGCACTCCCCAAACCCCTCCCTAATCCACATTCTTTCATTCCCATCTTTCCCCTCTCTGCTGCCCTCTTTACACTCTCATCACTTACCATCACTAACCACAATTTCTTCATGACTCTTGTTTTCTCTAC
ACCTGTCTTGTGACGGACTGAGATCCACGATTTCTTCACCCCTCCCCTTTCTCACTTCAACACCCTCTCCCATCTCTTTCTCCA
3103
STOP
rrj
jo
l<ei 5
ig. 3. Nucleotide sequence of the H L A - B 2 7 gene. Nucleotides are numbered from the Sac I site. The promotor ( T C T A A A ) and the polyadenylation
( A A T A A A ) signals are underlined. Splice sites and potential alternative splice signals are indicated by lines. The amino acids are placed above the triplets of
he exons in the three letter code.
Organization and Expression of the H L A - B 2 7 Gene • 375
Inspection of the H L A - B 2 7 gene sequence reveals several alternative
splice sites (AG/G) only a few of which do not result in stop codons. The
alternative splice acceptor sites described for the H - 2 K gene are found in
exactly the same positions in the H L A - B 2 7 gene (659 and 831, marked in
Fig. 3) together with the corresponding donor site in exon 2 at position 736.
The additional exon would code for 26 amino acids as in the H - 2 K gene. A
similar alternative transcript cannot be generated from the H L A - B 7 gene.
The presence of potential alternative splice signals in the H L A - B 2 7 gene
raises the question of whether they might be used. No c D N A clone coding
for H L A - B 2 7 has been described that is the product of an alternative
spliced transcript (16).
d
d
A complete c D N A sequence coding for the H L A - B 2 7 antigen (16) and
potential peptide sequencing data for this antigen (3) have been published
recently. We, therefore, do not discuss the protein sequence in detail. It is of
interest to note that the genomic sequence for the H L A - B 2 7 antigen
contains the triplet G C G at amino acid position 182 coding for alanine, as it
was found by protein sequencing (3). The c D N A clones, derived from a
homozygous B cell line, code for a valine ( G T G ) in this position. A second
amino acid replacement change is found in the leader at amino acid position
7 where the c D N A clone codes for a glutamine ( G A G ) , due to a change of
three nucleotides. Another additional nucleotide is found in the 3'untranslated region at position 3692. The insertion of a T destroys the Taq I site
found in the c D N A (16). This Taq I site is in so far important, because the
L G 2 line from which the c D N A clones were derived cannot contain the
polymorphic Taq I fragment of 3.5 kb. Its corresponding fragment must be
of 1800 bp. The five changes are the only differences found between the
c D N A sequence and the genomic D N A sequence. Since both genes code
for the same H L A - B 2 7 subtype (HLA-B27.1), we conclude that this might
be the first evidence for somatic mutations in B cell lines in a gene other
than immunoglobulin.
Comparison of the gene sequence of HLA-B27 with an allelic gene sequence. The comparison of the two H L A - B alleles, H L A - B 2 7 and the crossreactive H L A - B 7 , at the gene level shows a high degree of nucleotide
sequence conservation in both coding (exons) and non-coding regions
(introns), as well as in the flanking regions (Table 3). The highest level of
divergence is expressed in intron 1 and exon 1, which is not contained in the
mature H L A molecule. The degree of divergence is highest in the 5' half of
the gene (exon 1 to exon 3). The H L A - B 2 7 and H L A - B 7 genes are virtually
identical in the 3'part of the genes. Surprisingly, the exons and introns have
accumulated mutations to a similar degree.
The sequence comparison of exon 2 of the two alleles (Fig. 4) reveals that
most of the mutations are scattered. Only one cluster of changes is found to
the 3' end of this exon, from amino acid position 77 to 83.
COMPARISON:
Exon
1:
MET
HLA-B27
B
ARG V A L THR ARG PRO ARG THR L E U L E U L E U L E U L E U T R P G L Y ALA V A L A L A L E U THR G L U THR T R P ALA
ATG C G G G T C ACG GCG C C C CGA A C C C T C C T C C T G C T G C T C T G G GGG GCA G T G G C C C T G ACC GAG A C C T G G G C T
7
T
A2
T
GCC
G
T
C
G
A
C
C
C
T
C
C
C
C
G
H
X
X
Exon
2:
1
GLY
HLA-B27
10
SER
20
w
H I S S E R MET ARG T Y R P H E H I S THR S E R V A L S E R ARG PRO G L Y ARG G L Y G L U PRO ARG P H E I L E THR V A L
GGC T C C C A C T C C A T G AGG T A T T T C C A C A C C T C C G T G T C C CGG C C C GGC C G C GGG GAG C C C C G C T T C A T C A C C G T G
B7
T
A2
T
TT
A
y>
^
T A
^
G A
g
Z
30
GLY
HLA-B27
TYR VAL ASP
ASP
40
THR L E U P H E V A L ARG P H E ALA S E R A S P
50
^
GGC T A C G T G GAC GAC ACG C T G T T C G T G AGG T T C GAC AGC GAC G C C GCG AGT C C G AGA GAG GAG C C G CGG GCG C C G
O
B7
C
A
A2
Z
A
C
C
60
TRP
HLA-B27
I L E G L U G L N G L U G L Y PRO G L U T Y R T R P ASP
G AT
?
70
r
ARG
T G G ATA GAG CAG GAG GGG C C G GAG T A T T G G GAC CGG GAG ACA CAG A T C T G C AAG G C C AAG GCA CAG A C T GAC GCA
A C
A2
T
G
80
GLU
ASP
A
G
AA G T G
C
C
T
C
90
L E U ARG THR L E U L E U ARG T Y R T Y R ASN G L U S E R G L U A L A
GAG GAC C T G CGG A C C C T G C T C C G C T A C T A C AAC C A G AGC GAG G C C
B7
A2
A
ARG G L U THR G L N I L E C Y S L Y S ALA L Y S ALA G L U THR ASP
B7
HLA-B27
p
ALA ALA S E R PRO ARG G L U G L U P R O ARG ALA PRO
AG
T
A
G
G
G
G
G
Fig. 4. Sequence of the H L A - B 2 7 gene of exon 1 (leader) and exon 2 (1st domain) compared to equivalent regions in the H L A - B 7 (14) and H L A - A 2 (15)
genes. The amino acids are shown in the three letter code.
|
Organization and Expression of the H L A - B 2 7 Gene • 377
Discussion
We have shown that the cosmid clone CD2.6 contains one H L A class I
gene which is the H L A - B 2 7 gene. This human H L A gene can be expressed
in murine L cells and is recognized by monoclonal anti-HLA reagents even
when the heavy chain is combined with murine ß microglobulin. Several
monoclonal antibodies demonstrate that the epitopes, normally associated
with the H L A - B 2 7 molecule, are present on the surface of the transformed
L cells.
The D N A sequence and deduced amino acid sequence of the H L A - B 2 7
antigen were compared to the D N A sequence of its allele H L A - B 7 . As we
have discussed elsewhere (16), the polypeptide structure exhibits some
replacement changes unique to the H L A - B 2 7 antigen. Among them two
mutations are unusual: position 67 is occupied by a cysteine and position
131 by a serine. The change in position 131 generates a new restriction site,
G A G C T C , which is recognized by the enzyme Sacl. These changes could
be decisive in the peculiar role of H L A - B 2 7 as a risk factor for ankylosing
spondylitis.
Surprisingly, the comparison of the D N A structure of the H L A - B 2 7
gene with its allele H L A - B 7 revealed a high degree of homology between
the two genes. This is interesting in two aspects: Firstly, the HLA-B27/B7
pair is another example of human class I alleles showing a greater degree of
nucleotide conservation than allelic class I genes in the mouse (15, 29).
Thus, the two murine alleles H - 2 K and H - 2 K have a much lower level of
homology in the polymorphic exons 2 and 3 (10.3 and 11.2% divergence,
respectively), whereas the introns show the same degree of homology as the
human alleles (see Table 3). This higher mutation rate in the H - 2 K genes
could be due to their position centrometric to the I-A region, thus possibly
making it a preferred target for gene conversion events. This explanation is
supported by the sequences of H-2 class I genes other than H-2K. The
sequence comparison of the H - 2 D gene with its proposed allele H - 2 L (30)
( H - 2 D being less homologous) shows a level of divergence comparable to
human H L A - A and H L A - B alleles. Another explanation for the fact that
the human class I alleles are more similar than their murine homologues
could be based on the different evolution in the two species. It remains to be
explained why human class I alleles exhibit quite a high degree of divergence at the 5' half of the gene (exon 1 and intron 1).
The second aspect of interest concerns the differences found between the
H L A - B 2 7 and H L A - B 7 genes. The two H L A - B alleles are as homologous
as the H L A - A alleles and show to the 3'end of the gene, i.e. exon 4 to 3'
untranslated region, an even higher level of homology. This fact makes it
difficult to assign the cause for the association of H L A - B 2 7 with certain
diseases to particular structural features of the H L A - B 2 7 gene. Surprisingly, the highest degree of divergence is found in exon 1 and intron 2. The
leader peptide, encoded by exon 1, is cleaved off during maturation of the
2
b
b
d
d
d
378 • ELISABETH H . WEISS, W . K U O N , C . DÖRNER, M . L A N G , and G . RIETHMÜLLER
H L A class I molecule. The sequence of intron 2 is not part of the mRNA
and therefore not translated. Only in case of an alternative processed
transcript of the HLA-B27 gene involving intron 2 could this sequence be
of importance.
Several hypotheses have been proposed to explain the association of some
H L A alleles with certain diseases. The isolation and expression of the gene
encoding the H L A - B 2 7 antigen allows us to address the proposed mechanism with recombinant D N A techniques. Since we isolated the gene from a
so far healthy individual, we cannot rule out the highly unlikely possibility
that the H L A - B 2 7 gene of a patient might contain mutations. T R A P A N I et
al. (31) isolated a 3.5 kb Taq I fragment containing the 3'half of a supposed
H L A - B 2 7 gene from a spondylitic patient. The partial sequence of 180 bps,
derived from intron 6-exon 7-intron 7 presented in this report, is identical
to our sequence, except the G at position 3223 in intron 7 is lacking in their
sequence but is contained in the H L A - B 7 gene.
To examine the possibility that a putative «illness susceptibility» gene in
linkage disequilibrium with H L A - B 2 7 is responsible for the diseases associated with this antigen (the two-gene theory; see ref. 1 for review) we have
begun chromosomal walking experiments starting from the HLA-B27 gene,
in order to compare the organization of the H L A - B 2 7 haplotype with other
extended H L A - B loci.
The molecular mimicry hypothesis (the one-gene theory; see ref. 1 for
review) postulates that the molecular structure of infectious agents are
similar to those of the H L A antigens on the cell surface. In vitro
mutagenesis experiments may be helpful in defining HLA-B27 specific
epitopes that may cross-react with bacterial proteins. At present, we are
converting the H L A - B 2 7 unique replacement changes at amino acid positions 67 and 131 back to the «wildtype» configuration. Then we can
analyze whether these mutations affect the overall structure of the molecule
defined by monoclonal antibodies and human allogeneic cytotoxic T cells.
In summary, this paper describes the isolation of the H L A - B 2 7 gene
which, for the first time, allows the investigation of the mechanisms
underlying the observed association between HLA-B27 and various diseases.
Acknowledgements
K. BLÖMER is kindly acknowledged for her help in subcloning the two Pst I fragments. We
thank T . SCHLUNCK for the expert assistance w i t h the EPICS-analysis. Finally, we w o u l d like
to thank D r . D . SCHENDEL for reading the manuscript and for her useful comments and Mrs.
L . A . MONTANA for her patient secretarial help.
This w o r k was supported in part by the Deutsche Forschungsgemeinschaft (SFB 2 1 7 ) and in
part by the Genzentrum at the University of M u n i c h .
Organization and Expression of the H L A - B 2 7 Gene • 379
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a novel H - 2 K related m R N A . Proc. N a t l . Acad. Sei. USA 80: 7561.
WEISS, E., L . G O L D E N , R. ZAKUT, A . M E L L O R , K . FAHRNER, S. KVIST, and R. A.
FLAVELL. 1983. The D N A sequence of the H - 2 K gene: evidence for gene conversion a> a
mechanism for the generation of polymorphism in histocompatibility antigens. E M B C J .
3: 453.
MALOY, W . L . , and R. B. W A L L A C E . 1982. Primary structure of the H - 2 D alloantigcn.
Immunogenetics 16: 11.
TRAPANI, A . J . , C. A . NICKELSON, N . J . D E A C O N , D . J . H O O K E R , and I . F . C.
M C K E N Z I E . 1985. Molecular cloning and partial nucleotide sequence of a 3.5 kb H L \ B27-asociated fragment genomic D N A . Immunogenetics 22: 399.
d
29.
b
30.
31.
b
Dr. ELISABETH H . WEISS, Institute of Immunology, Goethestr. 31, D-8000 M ü n c h e n 2,
Federal Republic of Germany
Contents Volume 170 • 1985
Original Papers
BEJARANO, M . - T . , M . - G . MASUCCI, and E. K L E I N : Specific and Non-Specific Components in the Triggering of Proliferative and Cytotoxic Responses of T Lymphocytes
w i t h Different Cell Density
BETTENS, F., C. W A L K E R , G . D . BONNARD, and A . L . DE W E C K : Effect of Cyclosporin A
on the Early Activation of H u m a n T Helper Lymphocytes: Inhibition of R N A Synthesis and Modification of the Expression of Activation Antigens
BlANCHI, A . T . J . , L . M . HUSSAARTS-ODIJK, and R. B E N N E R : Secondary Delayed Type
Hypersensitivity to H - 2 Subregion-Coded Alloantigens
BÜSCHER, K . - H . , V . KLIMETZEK, and W . O P F E R K U C H : Influence of A n t i b o d y and
Complement Components on Phagocytosis and Chemiluminescence of Macrophages
GROENEVELD, P. H . P., T . E R I C H , and G . KRAAL: In Vivo Effects of LPS on B
Lymphocyte Subpopulations. Migration of Marginal Zone-Lymphocytes and I g D Blast Formation in the Mouse Spleen
G U E N I N , R., and C. H . SCHNEIDER: Studies on Monovalent Anaphylactogens: Evidence
for a Minimal Size of the Carbohydrate Auxiliary Group
HAUSTEIN, D . : Binding of DNP-Specific Receptor Material of N o r m a l Thymocytes to
DNP-Gelatin-Coated Dishes
H O D L E R , B., V . E V E Q U O Z , U . TRECHSEL, H . FLEISCH, and B. STADLER: Influence of
Vitamin D Metabolites on the Production of Interleukins 1, 2 and 3
HOROHOV, D . W . , R. N . M O O R E , and B . T . ROUSE: Herpes Simplex Virus-Specific
Lymphoproliferation: A n Analysis of the Involvement of Lymphocyte Subsets . . . .
H U M E , D . A . : Immunohistochemical Analysis of Murine Mononuclear Phagocytes that
Express Class I I M a j o r Histocompatibility Antigens
JEANNIN, J . - F . , D . REISSER, P. L A G A D E C , N . O . OLSSON, and F. MARTIN: Synergistic
Effect of Liposomes and Endotoxins on the Activation of Rat Macrophage Tumoricidal A c t i v i t y
JOSIMOVITS, O . , H . OSAWA, and T . DlAMANTSTEIN: The Mode of A c t i o n of the Calcium
Ionophore A23187 on T Cell Proliferation. I . The Ionophore Does not Replace
Lymphokines but Acts via Induction of I L - 2 Production on I L - 2 Responsive Cells . .
K ö L A R E , S., and G . SANDBERG: Studies on Thymocyte Subpopulations in Guinea Pigs.
V I . Differentiation of Precursor Cells in Vivo and in Vitro
L E P E - Z U N I G A , J . L . , J . S. Z i G L E R , jr., and I . G E R Y : Dual Effect of Phorbol Myristate
Acetate ( P M A ) on Murine Thymocyte Cultures
MÄNNEL, D . N . , W . D R Ö G E , and W . F A L K : A Combination of Soluble Helper Factors
Bypasses the Requirement for Stimulator Cells and Induces Nonspecific Cytotoxic T
Cell Responses
M A N D I , Y . , G . SEPRENYI, R. PUSZTAI, and I . B E L Ä D I : Are Granulocytes the Main
Effector Cells of Natural Cytotoxicity in Chickens?
MORISAKI, I . , S. KIMURA, M . T O R I I , S. M . M I C H A L E K , J . R. M C G H E E , N . OKAHASHI, and
S. HAMAD A: Cell Wall Preparation Consisting of Group A Carbohydrate and
Peptidoglycan Moieties from Streptococcus pyogenes Activates Murine B L y m p h o cytes
NIEDERWIESER, D . , D . FUCHS, A . HAUSEN, G . JUDMAIER, G . REIBNEGGER, H . W Ä C H TER, and C. HUBER: Neopterin as a N e w Biochemical Marker in the Clinical
Assessment of Ulcerative Colitis
N l H A S H I , Y . , Y . K O G A , H . GONDO, K . T A N I G U C H l , and K . NOMOTO: ThymusDependent Increase in N u m b e r of T Cells in Parathymic L y m p h Nodes Induced by
the Biscolaurine Alkaloid, Cepharanthine
3
175
434
192
390
402
412
158
256
460
381
211
164
338
327
146
284
293
320
351
476 • Contents
PATARROYO, M . , and M . JONDAL: Phorbol Ester-induced Adhesion (binding) among
H u m a n Mononuclear Leukocytes Requires Extracellular M g
and is Sensitive to
Protein Kinase C, Lipoxygenase, and ATPase Inhibitors
RAMADORJ, G . , F. TEDESCO, D . BITTER-SUERMANN, and K . H . M E Y E R ZUM BÜSCHENFELDE: Biosynthesis of the T h i r d (C3), Eighth (C8), and N i n t h (C9) Complement
Components by Guinea Pig Hepatocyte Primary Cultures
SANDBERG, G . , O . SÖDER, and J . TJERNBERG: Studies on Thymocyte Subpopulations in
Guinea Pigs, V I I . Characterization of Cell Populations Responsive to Guinea Pig
Interleukin 1 and Interleukin 2
SCHAAF-LAFONTAINE, N . , C. BALTHAZART, and R. J . H O O G H E : Membrane Carbohydrates of L y m p h o i d Cells: The Receptor for Interleukin 2
SETHI, K . K . , Y . OMATA, and H . BRANDIS: Contribution of Immune Interferon ( I F N - y )
in Lymphokine-Induced Anti-Toxoplasma A c t i v i t y : Studies with Recombinant
+ +
Murine I F N - Y
U L M E R , A . J . , W . SCHOLZ, M . ERNST, and H . - D . F L A D : Response of H u m a n T
Lymphocytes to Phytohemagglutinin ( P H A ) after Sequential Depletion of M o n o cytes, H L A - D R , L e u l l a , and L e u 7 Cells
VEERHUIS, R., L . A . VAN E S , and M . R. D A H A : In vivo Modulation of Rat Complement
Activities by Infusion of A n t i - H Antibodies
WEISS, E . H . , W . K U O N , C. DÖRNER, M . L A N G , and G . RIETHMÜLLER: Organization,
Sequence and Expression of the H L A - B 2 7 Gene: A Molecular Approach to Analyze
H L A and Disease Associations
ZAPF, S., and M . LOOS: Effect of E D T A and Citrate on the Functional Activity of the
First Component of Complement, C I , and the C l q Subcomponent
+
+
+
305
203
448
249
270
419
133
367
123
Short Communications
BESSLER, W . G . , B. SUHR, H . - J . BÜHRING, C. P. M U L L E R , K . - H . WIESMÜLLER, G .
BECKER, and G . JUNG: Specific Antibodies Elicited by Antigen Covalently Linked to a
Synthetic Adjuvant
WANGEL, A . G . , H . ARVILOMMI, and I . JOKINEN: The Effect of Phenytoin in vitro on
N o r m a l Mononuclear Cells and on H u m a n Lymphoblastoid B Cell Lines of Different
Ig Isotype Specificities
Abstracts: X V I I . Meeting of the Society of Immunology
239
232
.1-117
Authors' Index
ALHEID, U . 4 7
B O C K , S. 3 4
DEGWERT, J . 2 3
A L I , S. 3 2
BÖCK, G . 160
DEICHER, H . 3 7 , 4 7 , 1 6 9 , 1 8 3
ANDERER, F . A . 118
BÖCKER, W . 119
DEPPER, J . M . 84
ANDERSON, M . 4 5
BÖRNER, C . 7 7
DIAMANTSTEIN, T . 1 2 4 , 1 5 4 ,
ANDRIGHETTO, G . 1 9 2
BÖSING-SCHNEIDER, R . 143
ANTICA, M . 4 9
BÖTTGER, E . 5 8
DlBELIUS, A . 2 4
APFEL, H . 1
B Ö T T G E R , E . C . 12, 13, 14
DICKNEITE, G . 2 5
ARNOLD, B . 6 0
B O L T Z - N l T U L E S C U , G . 7, 5 9
DlEDRICHS, M . 2 6
ARVILOMMI, H . 2 3 2
BONNARD, G . D . 4 3 4
D I E R I C H , M . P . 105,138
AUSTEN, K . F . 131
BOSSLET, K . 15
D l E S F E L D , H . J . 1, 1 2 5
BRADE,H. 16,17
DIETRICH, H . 4 3 , 1 2 6
BACCARINI, M . 2 2 , 7 0
BRADE, L . 1 6 , 1 7
DIXON, F . J . 78
BALTHAZART, C . 2 4 9
BRADE, V . 191
DÖHRMANN, J . 5 7
BAMBERGER, U . 2 , 9 3
BRANDEIS, W . E. 9 2
DÖLKER, I . 7 2
B A R T L E Y , G . 131
BRANDIS, H . 2 7 0
DÖRNER, C . 88, 3 6 7
BARTSCH, H . 3 , 9 , 1 6 2
BRAUCH, H . 18
D O L D I , C . 41
BATSFORD, S. R . 1 5 7
BRAUN, R . W . 7 7
DOMZIG,W.27, 166,170
BAUM, H . P . 5 8
BREDA VRIESMANN, P . VAN
BAUM, W . 4 1 , 7 4
114
164
DREIKHAUSEN, U . 4 7
DRÖGE, W . 2 9 , 4 2 , 82,96,
BAUMGARTEN, H . 4 , 5 , 6 , 1 9 0
BRENDEL, W . 2 4 , 3 1 , 1 4 9
BAUMGARTNER, I . 7
BRETERNITZ, U . 1 2 5
109,146
BECHT, H . 3 5
BRÖCKER, E . - B . 193
E B E R L E J . 189
BECK, A . 163
BüHRlNG, H . J . 1 0 , 2 3 9
ECHTENACHER, B . 2 8
BECKER, G . 1 0 , 2 3 9
BUHL, R . 182
ECK, H . P. 29
BECKER, H . 8
BÜRKLE, C . 2 8
EHRET, W . 149
BEIN, G . 119
BÜSCHER, K . - H . 3 9 0
EICHMANN, K . 54, 6 5 , 9 7 ,
BEJARANO, M . - T . 1 7 5
BURGER, R . 9 5 , 1 2 5 , 1 3 5 , 1 8 2
BELÄDI, 1 . 2 8 4
BURKART, V . 161
EMMRICH, F . 3 0 , 1 8 6
BENNER, R . 1 9 2
BURMEISTER, G . 107, 1 9 3
ENDERS, G . 31
BERKOVIC, D . 9 , 1 6 2
BURMESTER, G . R . 1 9 , 6 4
ENGEMANN, R . 1 5 9
BERNDT, H . 155
BURMESTER, U . 3 4
EPPLEN,J.T. 32,54
BERTOVICH, M . 131
111,173,186
E R D E I , A . 105
BESSLER, W . 7 2 , 1 5 1
CAMPEN, T . J . 1 3 0
ERICH, T . 402
BESSLER, W . G . 10, 5 6 , 2 3 9
CANNISTRA,S.
ERNST, M . 4 1 9
BETTENS, F . 4 3 4
CARLS, C . 2 0
ERTEL, C . 9 , 1 6 2
B E T Z , M . 11, 1 7 7
C H A N G , H . - C . 113
Es, L . A . VAN 133
BEUSCHER, H . U . 191
CHIPLUNKAR, S. 21
EULITZ, M . 4 9
BIANCHI, A . T . J . 1 9 2
CIRSI, M . 9 8
EVEQUOZ, V . 2 5 6
BIESERT, L . 5 6
CRAMER, M . 108
51,52
BILLMANN, P . 163
FÄSSLER, R . 1 2 6
BlNNINGER, L . 80
D A H A , M . R . 133
FALK, W . 9 6 , 1 4 6
BIRK, G . 4 7
D A H R , \V. 18
FASSBENDER, B . 1 3 2
BlTTER-SUERMANN, D . 12, 13, D A M E R A U , B . 1 1 6 , 1 3 6 , 1 6 5
14, 5 8 , 1 0 4 , 2 0 3
DECKER, T . 22
FATHMAN,C. G . 4 0 , 1 7 5 , 1 7 6
FELGENHAUER, K . 168
N o r m a l numbers refer to abstract numbers of the abstract issue o f the X V I I . Tagung der
Gesellschaft für Immunologie, September 1985, V o l . 170, N o . 1/2. Bold-faced numbers refer
to page numbers o f original articles, V o l . 170, N o . 3 , N o . 4, and N o . 5.
4 7 8 • Authors' Index
FERBER, E . 110
HAMMER, D . K . 2
JOKINEN, I . 2 3 2
FIEDLER, F . 6 7
HARPPRECHT, J . 4 5
JONDAL, M . 3 0 5
FIEDLER, H . 3 3
HASLER, K . 163
JOSIMOVITS, O . 164
FINKBEINER, H . 3 4 , 1 5 5
HAUBECK, H . - D . 46
JUDMAIER, G . 3 2 0
F L A D , H . - D . 193,419
HAUSEN, A . 3 2 0
JÜRGENS, G . 160
FLEISCH, H . 2 5 6
HAUSTEIN, D . 1 6 4 , 1 5 8
JUNG,G. 10,239
FLEISCHER, B . 3 5 , 3 6
HEDERER, R . 28
FÖRSTER, O . 7 , 5 9
H E D R I C H , H . J . 141
FORBERG, K . 147
H E I N , R . 47
FRANKE, M . 118
H E I N L E , S. 5 6
FRANZ, A . 3 7
HEINZ, H . P. 48,101
KALIES, I . 79
FREUDENBERG, N . 112
HEISS, M . M . 4 9
KARAM, M . 125
FRICKE,M.169
HELMKE, K . 8
FRIEDRICH, W . 81
HEMMERLING, A . 5 0
KATUS, H . 8 6
KAUFMANN, S . H . E . 2 1 , 3 0 ,
FRÜHMARK, G . 148
HEMPELMANN, E . 1 2 5
FÜTTERER, A . 188
H E N F L I N G , M . 114
K E C K , K . 68
FUCHS, D . 3 2 0
H E R C E N D , T . 130
KELLER, R . 69
HERMANEK, P . 7 5
KERN,H.F.15
GÄRTNER, M . 129
HERRMANN, F . 5 1 , 5 2
KlDERLEN, A . F . 70
GASSMANN, W . 3 8
HESS, H . 103
KIESEL, U . 71,161
GATTNER, H . G . 6 8
HESSE, D . 165
G E H R I G , T . 121
H E U E R , J . 53
KIMURA, S. 2 9 3
K l R C H H O F F , H . 41
GEHRUNG, M . 93
HINTZ, P . 93
KIRCHNER, H . 4 1 , 1 6 7
GERY, I . 327
HOCHGESCHWENDER, U . 3 2 ,
KLAR, D . 50
GEUSENDAM,G.
119,155
KABELITZ, D . 117
K A L D E N J . R . 1 9 , 4 1 , 6 4 , 75,
76, 79
54,146
67,112
KLECH, H . 7
G I E D L , J . 75
HODLER, B . 256
K L E I N , E . 175
GLASSL,H.180
H O E G E N , P . VON 5 5
KLEINE, B . 72,151
GLEICHMANN, E . 6 2
HÖRL,W.163
KLIMETZEK, V . 3 9 0
G Ö H R I N G , P . 19
HÖVELS, A. 4 7
KLOSTERHALFEN, S. 73
GÖTTLINGER, H . 3 9
HOFFMANN, P . 5 6
KLOSTERHALFEN, W . 73
GÖTZE, 0 . 5 , 6 , 1 9 0
HOFFMANN, R . 5 7
KOCH, B . 74,75
GOLDMANN, S. F . 81
HOFFMANN, T . 1 3 , 1 4 , 5 8
K Ö L A R E , S. 3 3 8
G O N D O , H . 351
HOFFMANN-FEZER, G . 4 9 , 8 7
KöLBLE, K . 76, 79
GORONZY,J.40, 175,176
HOLZINGER, C . 7, 5 9
KÖLSCH, E . 2 3 , 4 6 , 5 3
GOTTSMANN, K . 137
HOOGHE, R . J . 249
KÖNIG, A . L . 77
GRACE, D . 34
H O R N , T . 14
KÖNIGSBERGER, H . 24
GRAMATZKI,M.41
HOROHOV, D . W . 4 6 0
KOFLER, R . 78
GREENE, W . C . 84,85
HORSTMANN, U . 6 0
KOGA, Y . 351
GRIFFINJ.51,52
H U A N G , J . - H . 113
KOHLEISEN, B . 79
GROENEVELD, P . H . P . 4 0 2
HUBER, C . 320
K O H L E R , H . 113
GUENIN, R . 412
HUNIG,T.61
K O L B , H . 71,161
GÜRTLER, L . 189
H U M E , D . A . 381
KOLSZYNSKI, M . VON 3 8
GUMPRECHT, H . 76
HURTENBACH, U . 6 2
KONICEK, K . 62
GUTEKUNST, R . 155
HUSSAARTS-ODIJK, L . M . 1 9 2
KOPONEN, M . 166
HUSSEY, R . E . 130
KORTMANN-HINNEBURG, C .
IMMELMANN, A . 6 3
KOWENZ, E . 134
HADAM,M.
129
183
HADDING,U. 13,14,104
KRAAL, G . 4 0 2
HÄCKER-SHAHIN, B . 42
HAMMERLING, G . J . 5 0 , 6 0
JABLONSKI-WESTRICH, D . 4 4
KRAFT, D . 7
HÄNSCH, G . M . 1 1 , 9 5 , 1 3 4 ,
JÄNCHEN, H . 169
KRAMER, M . 111
JAHN, B . 6 4
H A H N , H . 150
JANITSCHKE, K . 125
KRAMER, M . D . 8 0
K R AMMER, H . 6 3
HALA, K . 43
JANKOVIQD. 65
KRAMMER, P . H . 2 8
HAMADA, S. 2 9 3
JEANNIN,J.-F. 211
JOHNSON,J.P.26, 39,66,115
KRANZ, B . 4 9
177,185
HAMANN, A . 4 4
KRAPF, E . 9 2
A u t h o r s ' Index • 4 7 9
K R E T H , H . W . 81
MARTIN, R . 81
O T T E , G . 116
KRIEGBAUM, H . 8 2
MARTINEZ, J . 102
OTTE, M . 34
KRÖMER, G . 83
MASUCCI, M . - G . 175
KRÖNKE, M . 3 , 84, 8 5
M A U C K , J . 103
PAPE, G . R . 5 7
K R U G , M . 86
MAUER-GROSS, U . 104
KUMMER, U . 8 7
MELCHERS, F . 105
PATARROYO, M . 3 0 5
K U O N , W . 88, 3 6 7
MELCHERS, 1 . 6 5 , 9 7 , 1 0 6
PAUMGARTNER, G . 5 7
KURRLE, R . 166
M E O , T . 156
PEEST,D. 47,169
KYAS,U. 89,140
M E S K E , S . 163
PERMANETTER, W . 2 4
METZGER, S. 13
PETER, H . H . 1 2 0 , 1 2 9 , 1 6 3
M E U E R , S. 5 2 , 1 0 4
PETERHANS, E . 170
MEYER, J . 5
PFEFFER, K . 1 1 7
MEYER, T . F . 1 , 3 3 , 9 5
P F E I F L E J . 118
M E Y E R ZUM BÜSCHENFELDE,
PFIZENMAIER, K . 3 , 9 , 1 2 8 ,
LAGADEC, P . 211
LANDOLFO, S. 146
L A N G , B . 163
LANG, M . 3 6 7
LANGHORNE,J.21
142,162
K. H.203
P l C H L E R , W . J . 166
LASSMANN, H . 7
MICHALEK, S. M . 2 9 3
LAURELL, A . B . 48
MICHELS, E . 107
PlENNISCH, G . 2 0
POHL, C . 62
LEHLE, G . 90
MIERAU, R . 108
LEIBOLD, W . 4 1
M I H A T S C H , M . J . 157
POSCHMANN, A . 3 7 , 1 1 9
LEMM, G . 91
M I H M , S . 109
PUSZTAI, R . 2 8 4
LENHARD, V . 9 2
MINKENBERG,
LEPE-ZUNIGA,J. L . 327
M O H R , C . 76
LEONARD, W . J . 84
MOLL, H . I l l
L E V E N , J . P . 68
MOORE, R . N . 460
1.110
LIEBERKNECHT, L . 121
MORISAKI, 1.293
LÖGLER-ELLETT, G . 93
MOSSMANN, H . 2 , 9 3
LOERS, E . 8
M Ü G G E , K . 154
LOHMANN-MATTHES, M . - L .
22, 70
Loos, M . 4 8 , 1 0 1 , 1 2 3
LOPATTA, D . 192
MÜLLER, A . 137
MÜLLER, 1.112
M Ü L L E R - R U C H H O L T Z , W . 38,
45,69
L O V E T T , D . 8 9 , 94
MULLER, C . P . 1 0 , 2 3 9
Lucius, R . 1
MULLER, S. 113
LUDWIG, W . - D . 144
MYSLIWIETZ,J. 4 9
LÜBEN, G . 15
RAEDER, K . 168
RAMADORI, G . 2 0 3
RAMB-LINDHAUER, C . 1 2 0
RAUTERBERG, E . W . 2 0 , 86,
102,121,179,185
REDDIG, U . 34
REIBNEGGER, G . 3 2 0
REINHERZ, E . L . 130
REISSER,D.211
REITER, C . 188
R E N G E R , D . 169
RESCH, K . 89, 9 4 , 1 4 0 , 1 5 4
RESKE, K . 1 0 1 , 1 2 2 , 1 2 3
RESKE-KUNZ, A . B . 123,124
LÜHMANN, B . 165
NAGEL, G . A. 3,137,168
RiEBER, E . P . 57
LUMPP, E . 86
N A G E L K E R K E N , L . 114
RIETHMEISTER, G . 5 8
N E U , N . 43
RIETHMÜLLER, G . 3 9 , 5 7 , 8 8 ,
MA, D . L . 9 5
N E U M A N N - H A E F E L I N , D . 133
MACDERMOTT, R . P . 131
NIEDERWIESER, D . 3 2 0
M C G H E E J . R . 293
NIHASHI, Y . 351
R O E L C K E , D . 18, 7 7 , 1 3 4 , 1 8 1
MACHER, S. 121
NlXDORF, K . 145
RÖLLINGHOFF, M . 147
MÄNNEL, D . N . 9 6 , 1 4 6
N O L T E , R . 184
ROHWER, P . 74
MAIER, B . 9 7
NOMOTO, K . 3 5 1
ROLINK, A . 112
MAISCH, B . 9 8 , 9 9
N O O N A N , D . J . 78
ROTHER, K . 182
MALORNY, U . 1 0 7 , 1 9 3
NORDWIG, H . 115
ROTHER, U . 1 8 , 1 2 5 , 1 3 9 , 1 8 2
M A L Y , E . R . 100
NOWACK, H . 180
R O T T , R. 3 5
M A L Y , F . - E . 100
156, 1 8 8 , 1 8 9 , 3 6 7
RiTZ,J. 1 3 0 , 1 3 1
ROUSE, B . T . 4 6 0
MÄNDI, Y . 2 8 4
OPELZ, G . 92
R O Y E R , H . D . 130
MANKE, H . G . 9 2
OCHILEWSKI, U . 71
RUBIN, K . 48
M A N N , K . 148
OKAHASHI, N . 2 9 3
R Ü D E , E . 123, 1 2 4 , 1 3 2
MARQUARDT, P . 81
OLSSON, N . 0 . 2 1 1
R Ü H L , H . 144
MARTIN, F . 2 1 1
OMATA, Y . 2 7 0
RUKAVINA, V. 7 6
MARTIN, H . 101
OPFERKUCH, W . 3 9 0
RUMPOLD, H . 7
MARTIN, M . 9 4
OSAWA, H . 164
RUPPEL, A . 125
480 • Authors' Index
VEERHUIS, R . 133
SABBATH, K . 5 2
SIMON, H . G . 3 2
SANDBERG, G . 3 3 8 , 4 4 8
SIMON, M . M . 3 2 , 8 0 , 1 1 1 , 1 4 6 VITETTA, E . S. 85
SCHAAF-LAFONTAINE, N . 2 4 9
SÖDER, 0 . 4 4 8
SCHÄFER, H . 1 3 5
SOLBACH, W . 147
VOGT,A.
SCHANG, T . 1 5 9
SORG,C
V O G T , W . 1 1 6 , 1 3 6 , 165, 184
SCHAUENSTEIN, K . 8 3 , 1 2 6
SPAETH, E . 132
SCHAUER, U . 1 2 7
SPECHT, B . U . VON 2 4 , 1 4 8 ,
SCHEDEL, 1 . 4 7 , 1 6 9
V O G E L , L . 164
107,193
133,157
VORDERMEIER, H . - M . 5 6
149
SCHEINER, O . 7
SPERLING, U . 150
SCHELL, S. 145
SPETH, V . 110
WÄCHTER, H . 320
SCHENDEL, D . J . 2 6
S P Ö T T L , G . 148
WAGNER, H . 36
S C H E R B E R I C H , } . E . 103
SPRENGER, R . 151
W A H N , V . 139
SCHEUBER, P . H . 2
STACHEL, D . 189
WALDMANN, T . A . 85
SCHEURICH, P . 3 , 9 , 1 2 8 , 1 6 2
STADLER, B . 2 5 6
WALKER, C . 1 6 6 , 1 7 0 , 4 3 4
SCHIRRMACHER, V . 5 5 , 8 0
STÄB, F . 1 5 2
WALLACE, B . 54
S C H L E S I E R , M . 120, 129
STANLEY, K . K . 138
W A L T E R , M . 167
SCHLICK, E . 8 5
STEINER, H . 78
WALTER, P . 2 4 , 2 5
SCHLOSSMAN, S. F . 131
STELDERN, D . VON 124
WANGEL, A . G . 232
SCHMID, H . 6 4
STEVENS, R . L . 131
WANK, R . 2 6 , 6 6 , 1 1 5
SCHMIDT, R . E . 1 3 0 , 1 3 1
STOCKINGER, B . 153
WARNATZ, H . 91
SCHMIEDEL, D . 4 7
STOECK, M . 154
WASSNER, P . 68
SCHMITT, E . 1 3 2
STÖCKER, W . 3 4 , 1 1 9 , 1 5 5
W E B E R , T . 168
SCHNEIDER, C . H . 4 1 2
STOLPMANN, R . 1 5 0
W E C K , A . L . DE 1 6 6 , 4 3 4
SCHNEIDER, H . 133
STRIGL, G . 149
WEDEKING, U . 99
SCHÖNERMARK, S. 1 1 , 1 3 4
STROMAL, R . 78
WEHMEIER, U . 169
SCHOLLMEYER, P . 1 5 7
STRUVE, D . 3 4
WEHRMAKER, A . 2 9
SCHOLZ, W . 4 1 9
SUHR,B. 10,239
WEHRMANN, M . 4 7
SCHORLEMMER, H . U . 2 5
SUNDICK, R . S. 8 3
WEIDEMANN, M . J . 170
SCHRAMM, W . 189
S Z Ö T S , H . 156
W E I L A N D , E . 171
SCHREZENMEIER, H . 3 6
W E I L A N D , F . 171
SCHROD, L . 9 5 , 135
TANIGUCHI, K . 3 5 1
WEILER, E . 9 0 , 1 7 8
SCHROER, H . 1 3 6
TEDESCO, F . 2 0 3
WEISS, E . 8 8 , 1 5 6 , 1 7 2 , 3 6 7
SCHUFF-WERNER, P . 1 3 7 , 1 6 8
TEICHMANN, H . 144
W E I T Z E L , R . 122
SCHUH, R . 8 7
THAISS,F. 157
W E L T Z I E N , H . - U . 5 4 , 6 5 , 173
SCHULZ, T . 1 0 5 , 1 3 8
THEOFILOPOULOS, A . N . 7 8
W E N Z E L , B . E . 174
SCHULZE, M . 6 , 1 9 0
T H I E D E , A . 159
W E R N I C K E , D . 189
SCHWARZ, H . F . 15
THIEL, E . 49
W E S E L O H , G . 64
SCHWARZ, S. 126
THIELE, H . - G . 44
WESTPHAL, E . 45
SCHWERTZ, R . 1 3 9
THIERFELDER, S. 4 9 , 8 7
W E Y AND, C M . 4 0 , 1 7 5 , 1 7 6
SCHWINZER, B . 8 9 , 1 4 0
T H O E N E S , G . H . 158
W I C K , G . 4 3 , 78, 8 3 , 1 2 6 , 1 6 0 ,
180
SCHWINZER, R . 141, 154
TIMMERMANN, W . 159
SCRIBA,P. C . 1 5 5 , 1 7 4
TJERNBERG, J . 4 4 8
WIEGAND, R . 177
SEDLACEK, H . H . 1 5 , 2 5
TORII, M . 293
WIESMÜLLER, K . - H . 1 0 , 2 3 9
SEEMAYER, N . 100
T R A I L L , K . N . 160
W I L K E , J . 178
SEIDEL, J . 183
TRECHSEL, U . 2 5 6
WlLTSCHKE, C . 59
S E I T Z , C 119
T R E I C H E L , U . 161
WINGEN, A . - M . 121,179
SEITZ, R . - C . 3 7
TRENKMANN,J.
WINTER, U . 160
188
W O L F , G . 103
SELIGER, B . 142
WOLF, H . 78,180
SELINKA, H . - C . 143
Ü C E R , U . 3 , 9 , 128,162
SELS, F . 72
ULMER, A . J . 4 1 9
W O L F , M . 181
SEPRENYI, G . 2 8 4
ULRICHS, K . 6 9 , 1 5 9
WOLF, R . 1
SESSLER, M . 9 5
UMSCHEID, T . 158
WOLLENHAUPT, H . J . 139,
S I E B E R , G . 144
V A I T H , P . 163
WONIGEIT, K . 1 4 1 , 1 5 4
S l E G M U N D - S C H U L T Z E , N . 145
VALET, G . 87
WOODLAND, D . 6 5
182
SETHI, K . K . 2 7 0
Authors' Index
WOTTGE, H . U . 3 8
ZACHOVAL, R . 5 7
Z I E L A S E K J . 161
W U R L , M . 137
ZAPF, S. 123
ZIERZ, R . 6 , 1 9 0
Wussow,P. VON 183
ZEITZ, M . 144
Z i G L E R j . S . J r . 327
Z E N K E , G . 186
ZIMMERMANN, J . 191
Z I E G L E R - H E I T B R O C K , H . W.
ZÖLLER, M . 192
ZABERN, I . VON 184
ZACHARIOU, Z . 8 6 , 1 8 5
L . 187,188,189
ZWADLO, G . 193
Subject Index
Activation of macrophage tumoricidal
activity
Adhesion, p h o r b o l ester-induced . . . .
Adjuvant, synthetic
Alloantigens, H - 2 subregion
Anaphylactogens, carbohydrate auxiliary
group
Anaphylactogens, monovalent
Antigen, covalently linked
A n t i - H antibodies, complement activities
Anti-toxoplasma activity, I F N - y
ATPase inhibitors, adhesion
B lymphocyte subpopulations, effect of
LPS
B lymphocyte subpopulations, migration
B lymphocytes, streptococcus pyogenes
Biosynthesis of complement components, hepatocyte
Cl,effectofEDTA
C 1, functional activity
C l q , functional activity
Calcium Ionophore A 23187, T cell p r o l i feration
Carbohydrate auxiliary group, anaphylactogens
Carbohydrates, membrane
Cell density, T lymphocytes
Cepharantine, T cells
Chickens, natural cytotoxicity
Complement activities, a n t i - H antibodies
Complement components, C3, C8, and
C9
Complement components, phagocytosis
of macrophages
Complement, first component
Complement, i n vivo modulation . . . .
Cyclosporin A , effect on human T helper
lymphocytes
Cyclosporin A , RNA-synthesis
Cytotoxic responses, T lymphocytes . .
Cytotoxic T cell responses, helper factors
Cytotoxic T cell responses, nonspecific .
211
305
239
192
Cytotoxic T cell responses, stimulator
cells
Cytotoxicity in chickens, granulocytes .
412
412
239
Delayed type hypersensitivity, alloantigens
Delayed type hypersensitivity, secondary
DNP-gelatin-coated dishes, thymocytes
133
270
305
E D T A , functional activity of C I
Endotoxins, activation of rat macrophages
402
402
293
203
123
123
123
164
412
249
175
351
284
133
203
390
123
133
Gene, H L A - B 27
Granulocytes, effector cells of cytotoxicity
G r o u p A carbohydrate, streptococcus
pyogenes
Guinea pigs, thymocyte subpopulation .
Helper factors, cytotoxic T cell responses
Hepatocyte primary cultures, complement components
Herpes simplex virus, lymphocyte subsets
Herpes simplex virus, lymphoproliferation
Histocompatibility antigens, class I I . . .
H L A , disease associations
H L A , molecular approach
H L A - B 27, organization, sequence and
expression
I F N - y , recombinant
Ig isotype specificities, B cell lines . . . .
IgD-blast formation, mouse spleen . . .
I L - 2 production, ionophore A 23187 . .
I L - 2 responsive cells, calcium ionophore
A 23187
Immune interferon ( I F N - y ) , anti-toxoplasma activity
Interleukins 1, 2, and 3, vitamin D
metabolites
Interleukin 1 and interleukin 2, guinea
pig
Interleukin 2, lymphoid cells
Interleukin 2, receptor
Ionophore A 23187, I L - 2 production . .
146
284
192
192
158
123
211
367
284
293
338
146
203
460
460
381
367
367
367
270
232
402
164
164
270
3
434
434
175
146
146
256
448
249
249
164
484 • Subject Index
Liposomes, activation of rat macrophaLipoxygenase, adhesion
LPS, B lymphocyte subpopulations . . .
Lymphoblastoid B cell lines, effect of
phenytoin
Lymphoblastoid B cell lines, I g isotype
specificities
Lymphoid cells, membrane carbohydrates
Lymphoproliferation, herpes simplex
virus
Macrophage tumoricidal activity, liposomes and endotoxins
Macrophages, phagocytosis and chemiluminescence
Marginal zone-lymphocytes
Membrane carbohydrates, lymphoid
cells
Migration, B lymphocyte subpopulations
Mononuclear cells, effect of phenytoin .
Mononuclear leukocytes, phorbol esterinduced adhesion
Mononuclear phagocytes, class I I major
histocompatibility antigens
Murine mononuclear phagocytes, i m m u nohistochemical analysis
305
402
232
232
249
460
211
390
402
Phorbol myristate acetate, thymocyte
cultures
P M A , dual effect
Precursor cells, differentiation in vivo
and in vitro
Precursor cells, thymocyte subpopulations
Protein kinase C, adhesion
327
327
338
338
305
Rat complement
Receptor material, DNP-specific
Recombinant, I F N - y
RNA-synthesis, cyclosporin A
133
158
270
434
Streptococcus pyogenes, cell wall preparation
Synthetic adjuvant
293
239
T cell proliferation, calcium ionophore A
23187
T cells, cepharantine
T helper lymphocytes, cyclosporin A . .
T lymphocytes, depletion of monocytes
T lymphocytes, different cell density . .
T lymphocytes, proliferative and cytotoxic responses
T lymphocytes, response to P H A . . . .
T lymphocytes, triggering
Thymocyte cultures, effect of phorbol
myristate acetate
Thymocyte subpopulations, guinea
pigs
338,
Thymocyte subpopulations, interleukin 1
and interleukin 2
Thymocyte subpopulations, precursor
cells
Thymocytes, receptor material
Tumoricidal activity, macrophages . . . .
338
158
211
390
Ulcerative colitis, clinical assessment . . .
Ulcerative colitis, neopterin
320
320
232
305
Vitamin D metbolites, production of
interleukins 1, 2, and 3
256
249
402
232
305
381
381
Natural cytotoxicity, granulocytes . . . .
Neopterin, biochemical marker
284
320
Parathymic lymph nodes, cepharantine .
Peptidoglycan, streptococcus pyogenes .
P H A , T lymphocytes
Phagocytes, mononuclear
Phagocytosis, antibody and complement
Phenytoin, effect on normal human mononuclear cells
Phorbol ester-induced adhesion
351
293
419
381
164
351
434
419
175
175
419
175
327
448
448
3