Anti-MHC immunity detected prior to intentional alloimmunization

Cell fusion was performed between spleen cells from young BALB/cBy (H-2 ^d) mice which have never been immunized and SP2/0 mouse plasmacytoma cells. A monoclonal H-2-specific cytotoxic IgM antibody was obtained (By-1) which detected a new public biregional H-2 specificity, H-2.m210. The mcAb By-1 reacted strongly with H-2K^d, D^d, and H-2^s antigens, gave weak cross-reactions with H-2K^k, D^q, H-2^r, and H-2^v antigens and was negative with H-2^b, H-2^f, H-2^p, and H-2L^d antigens. A polymorphic reaction pattern was also observed on a panel of lymphocytes from B 10.W strains. The intriguing finding on this reaction pattern was the reactivity on H-2^d cells, including the syngeneic BALB/cBy and truly autologous cells. As shown by capping and immunoprecipitation experiments on H-2^d cells and by studies on H-2^d-transfected mouse L cells, the target molecules for McAb By-1 were H-2K^d and H-2D^d molecules. The BALB/cBy mouse, from whose spleen cells the McAb By-1 was obtained, survived after the fusion experiment, and serum was examined for the presence of cytotoxic H-2-specific antibodies during the rest of its life. At the time of the fusion, no autoreactive serum antibodies were found, but about 4 months later, we found in the serum of this mouse autoreactive H-2-specific cytotoxic IgM antibodies. The serum antibodies followed the same reaction pattern as that of the McAb By-1. As far as we know, this is the first report of autoreactive H-2-specific antibodies in serum of a mouse which has never been immunized and of the first natural autoreactive H-2-specific monoclonal antibody.Abbreviations McAb monoclonal antibody - MHC major histocompatibility complex - H-2 major histocompatibility complex of mice - CTLs cytotoxic T cells - FMF flow microfluorometry - FITC fluorescein isothiocyanate - LPS lipopolysaccharide W.E. coli 0111:134 - PBS phosphate-buffered saline - Iodogen 1,3,4,6,-tetrachloro-3,6-diphenylglycoluril - GAMIg goat-antimouse immunoglobulin - Staph-A Staphylococcus aureus Cowan I     

Fine specificity of cytotoxic T lymphocytes directed againstH-2L d

The fine specificity of cytotoxic T lymphocytes (CTL) directed againstH-2L ^d was analyzed by studying the lytic activity of BALB/cH- 2^dm2 (H-2L ^d loss mutant) anti-BALB/c-H-2 ^d CTL, generated in secondary mixed lymphocyte culture (MLC) against a panel of target cells of differentH-2 haplotypes. Target cells of allH-2 haplotypes tested, except that of the MLC responder, were lysed by anti-L^d CTL, although to a widely varying extent. The genes coding for antigens detected by anti-L ^d CTL were mapped to distinct regions in theH-2 ^d ,H- 2^dm1,H-2 ^q ,H-2 ^k , andH-2 ^b haplotypes. The sequence of lysis intensity against the variousH-2 haplotypes and theH-2 regions involved were as follows:L ^d ,D ^q L ^q ,D ^dm1 L^dm1,K ^k ,D ^b L ^b ,r, p, f, s, C3H.OH (K ^d D ^k L ^k ), strong lysis occurring againstL ^d and weak lysis againstH-2 ^s and C3H.OH.By monolayer adsorption and cold target inhibition experiments, it was shown that anti-L ^d CTL contained a CTL subset directed against a private L^d specificity, hitherto undetected by anti-L ^d antibodies. This subset of CTL was separate from the CTL subsets reacting againstH-2 ^q and against the mutant haplotypeH- 2^dm1. The reactions against the latter two haplotypes were also mediated by separate CTL subsets. It is concluded that the L^d molecule, to a varying extent, shares target antigens for CTL with K- and/or D-end H-2 molecules of all haplotypes tested. These antigens are detected by multiple subsets of anti-L ^d CTL. One CTL subset is directed against a target structure unique forL ^d (L^d private specificity).     

Fine specificity of a proliferating T-cell clone activated by a conformational determinant of the I-Ek molecule

We have examined the fine specificity of a stable Thy-1.2⁺, Lyt-1.2⁺, Lyt-2^–, and I-A^s– anti-I-E^k proliferating T-cell clone isolated from an A.TH anti-A.TL secondary mixed lymphocyte culture. Spleen cells from various I-A^k, E^k strains induced either a strong (A.TL, OH, and CBA) or a weak (AKR and B10.BR) proliferative response, although such cells expressed at their surface similar amounts of I-E^k antigens. Analysis of H-2 recombinant strains indicated that this clone recognized a conformational determinant carried by the E ^k E ^k dimer, but not on the Ea chain per se. Among the Fl hybrid strains in which the combinatorial E ^k E ^k product was detected by cellular binding with monoclonal E ^k -specific antibodies (mAb), some [(BIO.S(8R) × BlO.HTT) but not others (for example, B10.A(4R) × B10.A(5R)] were stimulatory. Seventeen anti-E^k mAb, regardless of the three spatially separated domains that they defined by antibody binding competition, completely inhibited the restimulation of this clone, whereas 15 other anti-A^k mAb failed to do so. This clone was not reactivated by stimulating cells from strains with the H-2 haplotypes p, j, v, b, r, and s but it proliferated strongly against cells from several H-2 ^d or H-2 ^q strains. Genetic evidence or blocking studies with selected mAb assigned these cross-reactive mixed lymphocyte reaction determinants to the A^d or A^q molecules, respectively. The data support the conclusion that alloreactive T cells may define a polymorphism of I-region coded products not detected by serological analyses and extend at the T-cell level the observations of serological cross-reactions between A and E molecules.     

H-2 haplotype specificity of molecular requirements for trinitrophenyl recognition by anti-hapten cytotoxic T lymphocytes

The requirement of direct covalent association of trinitrophenyl (Tnp) groups with cell surface components for functional interactions with anti-Tnp cytotoxic T lymphocytes (CTLs) was analyzed. The question was approached by comparing three different methods of modifying target cells with Tnp groups and analyzing the ability of three anti-Tnp effector populations with different H-2 haplotypes (H-2^k, H-2^d, and H-2^b) to lyse the syngeneic Tnp-modified cells. All effector cell populations were able to lyse in an H-2 restricted manner the appropriate target cell modified with 2,4,6-trinitrobenzenesulfonic acid. As previously shown, H-2^k anti-Tnp CTLs exhibited true H-2 restriction while H-2^d anti-Tnp and H-2^b anti-Tnp CTLs lysed the haptenated syngeneic target cell preferentially but not exclusively. Cells modified by either trintrophenylated bovine serum albumin (Tnp₃₅-BSA) or trinitrophenylated Sendai virus (Tnp-SV) were rendered susceptible to lysis depending upon the H-2 haplotypes of the target cells and the anti-Tnp effector cells. H-2^k anti-Tnp CTLs were able to lyse H-2^k target cells modified with either Tnp₃₅-BSA or Tnp-SV; however, H-2^d anti-Tnp or H-2^b anti-Tnp CTLS did not significantly lyse the H-2^d or H-2^b target cells modified by either Tnp₃₅-BSA or Tnp-SV. The results suggest that Tnp groups not covalently linked with cell surface specific components can be recognized by H-2^k anti-Tnp CTLs, but not by H-2^d or H-2^b anti-Tnp CTLs.     

Analysis of theD-region products ofH-2444using monoclonal antibodies reveals the expression of a new class I-like molecule

To analyze how many D-region-encoded molecules could be detected inH-2 ^q , we produced a panel of nine monoclonal antibodies from AKR (K^kD^k) anti-AKR.M (K^kD^q) immunizations. All of the D_q region antibodies cross-reacted on D^d and/or L^d, and all except one cross-reacted on D^b, confirming the previously observed serologic and amino acid sequence homology between theD-region products ofH-2 ^d ,H-2 ^b , andH-2 ^q . All of these monoclonal antibodies precipitated 46 000 dalton molecules from both cell-surface-labeled and biosynthetically labeled BIO.AKM spleen cells, indicating that all were reactive with class I-like molecules. Sequential immunoprecipitation analysis with one of these antibodies, 66-3-5, reveals the presence of a previously unidentified class I-like molecule. Tryptic peptide map analysis reveals that this molecule may be the product of a newly describedH-2D ^q -region gene.     

MHC recognition by clones of Mls specific T-lymphocytes

To test whether M1s determinants, like other non-MHC or nominal antigens, are recognized by T-cells in association with H-2 determinants, the in vitro proliferative responses of T-cell lines and clones were studied. Lines and clones were prepared by soft agar cloning (B10.BR x BALB/c)F₁ (H-2^k/H-2^d, M1s^b/M1s^b) T-cells responding in a primary MLR to AKD2F1 (H-2^k/H-2^d, M1s^a/M1s^a) stimulator cells. All the T-cell clones obtained could respond equally well in a proliferative assay to the Mls^a determinant in association with the H-2 haplotype of either parent, i. e., DBA/2 (H-2^d, M1s^a), and AKR (H-2^k, M1s^a) both stimulated equally well. When the T-cell lines and clones were screened against stimulators from recombinant inbred (RI) strains, it became apparent that strains exhibiting the H-2^b, M1s^a genotype stimulated poorly or not at all. This shows that the T-cell response to M1s^a involves MHC recognition, and raises the possibility that the response to M1s^a can involve recognition of H-2 specificities shared between the H-2 ^k and H-2 ^d haplotypes.Abbreviations used in this paper MHC major histocompatibility complex - MLC mixed lymphocyte culture - IL-2 interleukin 2 - Con A concanavalin A - RI recombinant inbred Howard Hughes Medical Institute     

A third class I major histocompatibility complex antigen encoded by a gene in the D region of the H-2 d haplotype recognized by cytotoxic T lymphocytes

The D region of the H-2 ^d haplotype contains five class I genes: H-2D ^d , D2 ^d , D3 ^d , D4 ^d and H-2L ^d . Although previous studies have suggested the presence of D-end encoded class I molecules in addition to H-2D^d and H-2L^d, segregation of genes encoding such molecules has not been demonstrated. In this report we have used cãtotoxic T lymphocytes (CTL) to examine the D region of the H-2 ^d haplotype for the presence of additional class I molecules. CTL generated in (C3H × B6.K1)F₁ (K ^k D ^k , K ^b D ^b ) mice against the hybrid class I gene product Q10^d/L^d expressed on L cells cross-react with H-2L^d but not H-2D^d molecules, as determined by lysis of transfected cells expressing H-2L^d but not H-2D^d. Although H-2L^d-specific monoclonal antibodies (mAb) completely inhibit H-2L^d-specific CTL from killing B10.A(3R) (K ^b D ^d L ^d ) target cells, only partial inhibition of anti-Q10 CTL-mediated lysis was observed, suggesting the presence of an additional D-end molecule as a target for these latter CTL. To identify the region containing the gene encoding the Q10 cross-reactive molecule, we show that anti-Q10 CTL lyse target cells from a D-region recombinant strain B10.RQDB, which has H-2D ^d , D2 ^d , D3 ^d , D4 ^d , and H-2D ^b but not the H-2L ^d H-2 ^d , and H-2L ^d (including D2 ^d , D3 ^d , and D4 ^d , lacks this anti-Q10 CTL target molecule. Together, these data demonstrate that a class I gene mapping between H-2D ^d and H-2L ^d encodes an antigen recognozed by anti-Q10 CTL. A likely candidate for this gene is D2 ^d , D3 ^d or D4 ^d .     

The spatial relationship of the viral and H-2 antigens recognized by anti-viral CTLs

With the use of monospecific rabbit anti-G protein and mouse monoclonal anti-H-2K^k, we have analyzed the spatial relationship of the serologically defined H-2K^k antigens and the major surface glycoprotein (G protein) of vesicular stomatitis virus (VSV) to those antigens recognized by B10.A (k, d) anti-VSV cytotoxic T lymphocytes (CTLs). The ability of monoclonal anti-H-2K^k or rabbit anti-G protein to inhibit specifically the cytolytic activity of B10.A anti-VSV CTLs indicates that the G protein and the H-2K^k molecules are in close proximity to the viral and H-2K^k antigens recognized by the anti-VSV (CTLs. By the method of sequential immunoprecipitation, we also demonstrated that only 10–30 percent of the serologically defined G and H-2K^k molecules are in theG-H-2K ^k complexes.Abbreviations used in this paper Con A Concanavalin A - cpm counts per minure - CTLs cytotoxic T lymphocytes - E: T ratio effector: target ratio - G major surface glycoprotein of VSV - MHC major histocompatibility complex - MOI multiplicity of infection - NP40 Nonidet-P40 - PAGE polyacrylamide gel electrophoresis - PBS phosphate-buffered saline - SaCI Staphylococcus aureus, Cowan I strain - SDS sodium dodecyl sulfate - UV ultraviolet light     

Receptor specificity,functional characteristics,and cell-surface phenotype of a highly selected anti-I-Ab-specific,long-term T-cell line

A highly selected alloreactive T-cell line was developed by repeated restimulation of B10.D2/n lymph-node cells with irradiated C57BL/10Sn (BIO) spleen cells in long-term MLC for up to 2 1/2 years. Continuous growth of the line requires restimulation every 2 to 4 weeks with fresh H-2^b stimulator cells. The line proliferates strongly against H-2^b but not againstH-2 ^d ,H-2 ^f ,H-2 ^q ,H-2 ^r , orH-2 ^s stimulators. Analysis of recombinant mouse strains showed that the proliferative response is directed against I-A^b but not K^b or D^b determinants. During the growth period of the line, strong cross-reactivity with H-2^p (B10.P) and weak cross-reactivity with H-2^k strains (e.g., CBA/J and B10.BR) was observed. A clone with exquisite specificity for I-A^b, but with no cross-reactivity with H-2^p or H-2^k was isolated from the line; thus clonal heterogeneity of the line still exists despite the highly selective growth conditions. — The majority of T cells from the line or clone were shown to bind I-A^b but not K^b or D^b determinants either spontaneously during restimulation with fresh B10 stimulator cells or via membrane vesicles expressing I-A^b determinants. No killing activity by the line in either specific or nonspecific cytolytic T-cell assays was observed nor was the T 145 glycoprotein, characteristic of killer T cells, detected.Abbreviations used in this paper B6 C57BL/6J - B10 C57BL/10Sn - Con A Concanavalin A - CTL cytotoxic T lymphocyte - FCS fetal calf serum - FDA fluorescein diacetate - FITC fluorescein isothiocyanate - Ia I-region-associated antigens - LPS lipopolysaccharide fromE. coli - Lyt T-lymphocyte-defined antigen - MLC mixed leukocyte culture - NP-40 nonidet P-40 - PAGE pofyacrylamide gel electrophoresis - PHA phytohemagglutinin fromPhaseolus vulgaris - PM plasma membrane - SDS sodium dodecyl sulfate - TCGF T-cell growth factor(s) - TdR thymidine     

H-2-linked genes determine the level of the primary in vitro anti-Mls response

The level of cell proliferation and interleukin-2 (IL-2) production observed in an anti-Mls mixed lymphocyte reaction between spleen cells from H-2 compatible, Mls incompatible mouse strains is determined by the H-2 haplotype of the mouse combination. Thus, while AKR (H-2 ^k) spleen cells stimulated strong M1s^a responses in H-2^k responder cells, AKR H-2b spleen cells stimulated no or negligible M1s^a responses in responder cells from H-2 ^bmouse strains. This effect was observed at the levels of IL-2 production and cell proliferation. The magnitude of the response observed using F₁ (H-2 ^k/H-2 ^b) responder cells was found to be a function of stimulator rather than responder cells. The poor stimulatory capacity of AKRH-2 ^bspleen cells was also shown not to be due to the loss of the stimulatory Mls ^aallele during the construction of the congenic strain from AKR and C57BL/6 parental strains. Using stimulator cells from a second series of congenic mice, we found H-2 ^b(strain DLLP) again to represent a poorly Mls^a stimulatory H-2 haplotype. In addition, H-2^q (DBA/1) cells displayed very poor Mls^a stimulatory potential while H-2^d (D1.C) cells were efficient Mls^a stimulators. Again the effect was shown to be at the level of the stimulator cells. In toto, our findings indicate that the H-2 ^kand H-2 ^dhaplotypes encode strong Mls^a stimulatory potential while the H-2 ^band H-2 ^qhaplotypes determine poor Mls^a stimulatory potential in primary in vitro responses, measured as cell proliferation and IL-2 production.Abbreviations used in this paper: CTL cytotoxic T lymphocyte - IL-1 interleukin-1 - IL-2 interleukin-2 - MLR mixed lymphocyte reaction - NMS normal mouse serum     

Identification of specificityH-2.7 as an erythrocyte antigen: Control by an independent locus,H-2G,between theS andD regions

SpecificityH-2.7 is expressed predominantly on erythrocytes and controlled by a gene that maps within theH-2 gene complex at a locus, designated asH-2G, which apparently lies between regionsS andD. Three phenotypes have been observed with respect to this antigen: a) positive by direct test and absorption (haplotypesH-2 ^f ,H-2 ^j ,H-2 ^p ,H-2 ^s ); b) positive only by absorption (H-2 ^k ); and c) negative (H-2 ^b ,H-2 ^d ,H-2 ^q ). New crossover positions have been established for severalH-2 recombinants based on classifications for theH-2G locus.     

A novel MHC class I-related molecule expressed on mouse thymic stroma cells and mature lymphocytes

A monoclonal antibody (mAb) TP-3 has been established by immunizing rats with the BALB/c mouse thymic epithelial cell line TEL-2. The TP-3 antigen is expressed on stroma cells of thymus, spleen, and lymph node in syngeneic BALB/c mice (H-2 ^d ). This antigen is also expressed at a low level on the cell surface of immature thymocytes, and at a high level on mature T and B cells. In allogeneic mice such as C57BL/6 (H-2 ^b ) or C3H (H-2 ^k ), no cells expressed the TP-3 antigen. Using H-2 congenic mice, reactivity with mAb TP-3 was found to map to a region of H-2D ^d L ^d or between D ^d and Qa, suggesting that TP-3 is a major histocompatibility complex (MHC) class I antigen. However, immunoprecipitation analysis indicated that this antigen is not identical to the classical mouse class I molecules in terms of molecular size, antigenicity, and tissue distribution.     

Murine cytotoxic T-cell response to alphavirus is associated mainly withH- 2D k

A secondary in vitro response to alphaviruses Bebaru, Sindbis, and Semliki Forest is described. Optimum response appears at day 5–6 of culture. The cells responsible for lytic activity are nonadherent, -positive, Ig^–, and mainly Ly-2.1 positive. Out of five haplotypes tested (H- 2 ^d ,H- 2 ^b ,H- 2 ^s ,H- 2 ^q , andH- 2 ^k ) onlyH- 2 ^k was a responder. Genetic mapping of the response located it solely in theD region of theH- 2 complex. The other four haplotypes responded with a high antiself activity after a second stimulation with viruses. This antiself response also maps in theD region of theH- 2 complex. No complementation was observed in F₁ hybrids between responder and nonresponder strains.     

Serological characterization of previously unknown H-2 molecules identified in the products of theK d andD k region

The molecular relationship between H-2 private and some public specificities was studied in C3H.OH (H-2 ⁰² ) mice using surface-antigen re-distribution methods. Besides the K^d- and D^k-region antigens, which can be capped by antisera against the private and public specificities characteristic for a given allele, a previously unknown type of molecule was found in the products of both theK ^d andD ^k regions. These can be capped by the respective anti-private serum but not by antisera against some public specificities. The two K^d-region molecules are provisionally named H-2K1^d and H-2K2^d. We detected them onH-2 ⁰² (K ^d ,I ^d ,S ^d ,D ^k ) and also onH-2 ^dx (K ^d ,I ^f ,S ^f ,D ^dx ) T lymphocytes. Similarly, the two types of molecules detected on the products of theD ^k region are provisionally named H-2D1^k and H-2D2^k. The serological characteristics of these molecules are described. When compared with the products of theD ^d region, in which we previously described three different molecules (H-2D^d, H-2M^d, and H-2L^d), the mutual relationship between H-2K1^d and H-2K2^d as well as between H-2D1^k and H-2D2^k appears to be similar to that between H-2D^d and H-2M^d. In the absence of relevant recombinants or informative biochemical data, it is, however, difficult to establish homology between molecules produced by differentK- andD-region alleles.     

Paired H-2-specific monoclonal antibodies react differently to red cells and lymphocytes

Two hybrid clones from a fusion of C57BL/6 anti-DBA/2 spleen cells and the myeloma line Sp2/0 secrete antibodies reactive with a product of the murine major histocompatibility complex (MHC). The two antibodies are provisionally designated 513.11 and 513.29. Both react in rabbit-complementmediated cytotoxicity with spleen cells of H-2 ^d , H-2 ^f , H-2 ^r , and H-2 ^p strains. In addition, both antibodies hemagglutinate red blood cells from these strains. S13.11 is also cytotoxic for H-2 ^a , H-2 ^k , H-2 ^u , and H-2 ^v spleen cells but does not hemagglutinate red blood cells from mice bearing these haplotypes. With the exception of H-2 ^v , this strain pattern mimics the public specificity H-2.8. Quantitative absorption of S13.11 shows that H-2 ^d cells are twice as efficient as H-2 ^k cells in their ability to remove the 513.11 antibody. 513.29 reacts weakly in cytotoxicity with H-2 ^kv spleen cells and does not react with cells from H-2 ^u or H-2 ^v . Blocking studies indicate that 513.11 and S13.29 react with the same or a closely related molecule on the cell surface.     

Resistance to cell-mediated cytotoxicity is correlated with reduction of H-2K gene products in AKR leukemia

AKR leukemia cell lines differing in the amount of H-2K and H-2D antigens expressed on the cell surface were used to assess cell-mediated immune responses in syngeneic mice against Gross/AKR murine leukemia virus (MuLV)-induced tumors. Leukemic cells with reduced expression of H-2K^k antigens were inactive as inducers of Gross-MuLV/H-2^k-specific cytotoxic T lymphocytes (CTL) and resistant to lysis by CTL raised against H-2K^k positive AKR leukemia cells. H-2K^k positive leukemias induced cytotoxic effectors, which upon restimulation in vitro, lysed the stimulating and other H-2K^k positive leukemia cells. In antibody inhibition experiments, T-cell-mediated cytotoxicity to these leukemias could only be inhibited by antisera and monoclonal antibodies specific for the H-2K^k antigens. Due to this specific role of H-2K^k antigens in T-cell cytotoxicity to Gross/AKR MuLV-induced tumors, reduced expression of H-2K^k antigens on spontaneous AKR leukemic cells could have important implications for surveillance of these neoplastic cells.Abbreviations used in this paper CTL cytotoxic T lymphocytes - MuLV murine leukemia virus     

Lateral diffusion and patch formation of H-2Kk antigens on mouse spleen lymphocytes

We have studied the diffusion and aggregation of H-2K^k antigens labeled with a fluorescent anti-H-2K^k monoclonal antibody (IgG) on mouse splenic lymphocytes, employing fluorescence photobleaching recovery and fluorescence microscopy. The H-2K^k antigens were initially distributed homogeneously on all lymphocytes. Upon antibody binding, sub-micron patches were formed on 50–60% of the cells. A lateral diffusion coefficient, D, of 7.1·10^?10 cm²/s and a mobile fraction of 0.73 were found for H-2K^k antigens on diffusely-labeled cells, while these antigens were immobile (D?5·10^?12 cm²/s) on patched cells. The patched and nonpatched sub-populations did not correspond to B- and T-lymphocytes. Subjection to low temperature or treatment with NaN₃ or cytoskeleton-disrupting drugs did not affect the diffusion or patching of H-2K^k, indicating no involvement of metabolic energy or drug-sensitive cytoskeletal components. These findings could be related to the interactions of H-2 antigens on the cell surface, and to the different susceptibilities of various cells to lysis by cytotoxic T-cells.     

Polymorphism of Hh-1, the mouse hemopoietic histocompatibility locus

The major goal of these studies is to more fully assess the polymorphism of the hemopoietic histocompatibility (Hh) genetic system. H-2 homozygosity is required for optimal immunogenicity of bone marrow cell (BMC) grafts, and hybrid resistance to grafts of parental strain BMC by irradiated H-2 heterozygous F₁ hybrid mice suggests that Hh-1 antigens are inherited recessively. The Hh-1 antigens are also expressed on other normal hematopoietic cells and lymphoid tumors, and natural killer cells are the effectors which mediate the elimination of BMC grafts in an Hh-specific manner. Previous studies have demonstrated three different antigens mapping to the Hh-1 locus near H-2D. We test the expression of Hh-1 on BMC of all nonrecombinant H-2 haplotypes of independent origin and H-2 ^j , a presumed natural recombinant. Hh-1 typing is based on the pattern of growth and rejection in a panel of hosts. F₁ hybrids with H-2 ^b , H-2 ^d , and H-2 ^k are produced and used as donors and hosts to confirm the phenotype. Grafts of b-, d-, and j-haplotype marrow serve as prototypical examples of determinants that are provisionally designated as 1, 2, and 3, respectively. We describe a new determinant, 4, in the k haplotype. It is non-codominantly expressed, maps to H-2D, and is also expressed on H-2^b BMC. NZW, H-2^Z grafts exhibit a phenotype similar to k, but express a unique determinant 5 which can be distinguished from determinant 4. This additional determinant is also expressed by the b haplotype. The d, f, and p haplotypes all express determinant 2, and grafts of j-haplotype marrow are found to express determinants 2 and 5 in addition to determinant 3. The q and r haplotypes are null for all known determinants. Finally, we describe a phenotype which is a new combination of previously described determinants: s-haplotype grafts express determinants 1, 2, and 4. The polymorphism of Hh-1 detected thus far consists of seven alleles which are combinations of five distinct determinants.     

Target-cell recognition by cloned lines of influenza virus-specific cytotoxic T lymphocytes. Selective inhibition by a monoclonal H-2-specific antibody

A monoclonal H-2^d-specific antibody markedly inhibits target-cell lysis mediated by two influenza virus A/JAP/57-specific, H-2K ^d -restricted cloned CTL lines. Three other A/JAP/57-specific, H-2 ^d -restricted CTL clones (two of which are also restricted to H-2K ^d in target-cell recognition) are only minimally inhibited by this monoclonal antibody. The inhibitory effect of the antibody is not due to selective binding to certain cloned CTL lines but rather is due to blocking of a determinant on the target cell. The monoclonal antibody produces partial inhibition of lysis mediated by a heterogeneous population of A/JAP/57-specific, H-2 ^d -restricted CTL. Likewise the profound, selective inhibition of cytolysis produced by the H-2^d-specific monoclonal antibody could not be reproduced with a conventional H-2^d alloantiserum. These observations suggest that more than one site on a particular H-2K or H-2D molecule can serve as a determinant for H-2-restricted CTL recognition. They furthermore imply that there is more than one recognition structure (receptor) for self MHC products clonally distributed among a population of H-2-restricted CTL directed to a particular antigen.     

Genetic control of immune responses of inbred mice: Responses against terpolymers poly(glu57lys38ala5) and poly(glu54lys36ala10)

The immune response patterns of inbred and congenic strains of mice against terpolymers poly(glu⁵⁷lys³⁸ala⁵) and poly(glu⁵⁴lys³⁶ala¹⁰) have been studied. Initial recognition of the polymers is ascribed to ‘GA’ receptors (Ir-GA gene product) on T cells of mice ofH-2 haplotypes,a,b,f,k ands, and ‘GL’ receptors (Ir-GL gene product) of mice ofH-2 ^p,H-2 ^q andH-2 ^j haplotypes, and to GA and/or GL receptors of mice ofH- 2^d andH- 2^r haplotypes. The specificity of the antibody is directed predominantly against GL. The inability to elicit antibody with GA specificity has been ascribed to the lack of significant concentrations of GA sequences in the polymers to interact with appropriate receptors on B cells. The weakest responders were mice of H-2^b haplotype. F¹ hybrids (responders×nonresponders) were all responders demonstrating the dominant character of responsiveness. Wide variations in antibody levels produced among strains of mice of theH-2 ^k andH-2 ^b haplotypes are ascribed to genes not linked toH-2.