Tissue graft rejection in mice

A liver-slice to kidney-bed grafting system was used to study the course of rejection of a specific tissue across various genetic barriers in inbred strains of mice. Rejection or survival, scored histologically at various times after grafting, demonstrated that multiple nonH-2 differences cause rejection at least as rapidly asH-2 differences. Differences at theK end of the mouse major histocompatibility complex cause tissue rejection more rapidly than do differences at theD end of the complex. The latter differences cause chronic rejection similar to that found across several minorH locus barriers. TheH-2 haplotype carried by the recipient or the strength of theH-2 antigens of the donor affect the survival time in liver tissue grafts. Studies employing this model system will contribute to the definition of different immunogenetic parameters affecting survival of various tissues in a genetically well-defined animal model.     

Tissue graft rejection in mice

A tissue slice-to-kidney bed grafting system is used to study the mechanism of specific tissue rejection (in this case, rejection of liver tissue) over a series of histocompatibility barriers other than the H-2 barrier. Using the method described, it is possible to obtain a pattern or time-course picture of the immunological process, rather than a mean survival time. It is clear from histological observations of these patterns that, although there are considerable differences in numbers of liver grafts which survive for long period's across the several histocompatibility barriers studied, some grafts in almost every case survive the immunological challenge elicited by the genetic barriers. Grafts of liver tissue are therefore similar, but not identical, in survival patterns to grafts of tumor, ovary, and skin. These studies also indicate that immunological mechanisms controlling rejection of tissue over H barriers other than H-2 differ from those controlling rejection over the major histocompatibility barrier in the mouse.     

H-2 and Background influences on tissue grafts across the H-Y barrier

Male liver was grafted to kidney beds in syngeneic female mice. Relative influences ofH-2 haplotype, genetic background or interaction ofH-2 haplotype with genetic background on anti-H-Y response were evaluated using 27 inbred strains carrying eightH-2 haplotypes of independent origin and three naturally occurring recombinants. Females ofH-2 ^b haplotype acutely rejected the male graft as is reported for other tissue graft systems. AnH-2 haplotype influence was found for all haplotypes studied, with a greater variation of immunologic response revealed by histological analysis of liver grafts than is demonstrated by skin grafts. Strains carryingH-2 ^k ,H-2 ^j andH-2 ^p haplotypes expressed the greatest range of immunological variability with responses ranging from graft proliferation to graft rejection. Strains carrying theH-2 ^d haplotype had the most consistent responses with little reaction to the graft. The strong immune response by SJL/J (H-2 ^s ) female mice to the H-Y antigen is not typical of otherH-2 ^s strains, but is compatible with the reported hyperresponsiveness of this strain to alloantigens.     

Host response to tissues placed in the anterior chamber of the eye: demonstration of migration inhibition factor and serum blocking activity.

Allograft immunity of rats to transplantation antigens was demonstrated by in vitro migration inhibition procedures. Spleen cell suspensions from rats sensitized to histocompatibility antigens by skin graft or anterior chamber implants failed to migrate normally in vitro when incubated in the presence of appropriate donor tissue extracts. Tissue grafts transplanted across both major and minor histocompatibility barriers had prolonged survival within the anterior chamber. However, 2 weeks after implantation, the recipient rats showed detectable sensitization to their implants. Serum from 14- to 32-day implant-bearing rats blocked the migration inhibition found in the implanted rats in the presence of corresponding tissue antigen. Such blocking activity was undetectable in the serum from rats which had implants for 7 days or after rejection of the implant was evident. MIF production in implanted and skin-grafted rats was evident at significantly different times in relation to the graft rejection. The asynchrony of MIF synthesis observed in these experimental animals leads us to postulate that the graft rejection and MIF production may be mediated by distinct lymphocyte populations. In addition, the route of the antigen presentation may account in part for the observed differences.     

The histocompatibility-2 system in wild mice

The I-region gene products of 29 wild-derivedH-2 haplotypes on a B10 background (B10.W congenic lines) were typed with alloantisera which detect 17 inbred I-region antigens. Five new I-region antigens were defined by expanding the inbred line panel ofH-2 haplotypes to includeH-2 ^u , H-2^v, andH-2 ^j . Based on serological analyses of the inbred and B10.W lines, the polymorphism of theIA gene (or genes) is estimated to be at a minimum of 15 alleles and theIE gene (or genes) at a minimum of 4 alleles. These results indicate that theIA subregion is more polymorphic than theIE subregion. By combining the I-region typing data with theH-2K andH-2D region typing data reported previously, a total of 11 new natural recombinants of inbredH-2 alleles were detected among the B10.W lines.     

The effect of H-21J disparity on induction of allotransplantation tolerance by Lentil Lectin

The effect of an H-21J disparity on skin graft survival was studied in 18 mouse donor-recipient strain combinations, in which the recipients were treated with an efficient immunosuppressant, lentil lectin (LCA). The simultaneous I-J disparity essentially had no (or a slightly adverse) effect on the graft survival times in strain combinations differing at the K and I-A loci or in the entire H-2 complex. In two strain combinations incompatible at the D locus, the simultaneous I-J disparity promoted graft survival. The disparity at the I-J locus therefore seems to have only a marginal effect on the survival of allografts in most of the LCA-treated recipients, but it may promote graft survival in some animals. A similar tolerance-promoting effect was also observed with D disparity.     

A histocompatibility region (H-2IC) in theIC region of theH-2 complex of the mouse

Skin graft rejection in congenic pairs of mice differing only at theH-2 complex appears to be influenced by at least 3 genes (H-2K, H-2D, H-2I); we now describe a fourth,H- 2IC: Grafts transplanted across anIC difference are sometimes rejected. TheI-C regions of three differentH-2 haplotypes (d,k,s) were studied in different combinations, and variable patterns emerged: (a)IC ^d : B10.S(7R) show delayed or no rejection of first B10.S(9R) grafts, but grafts to immunized recipients were usually rejected in 20 days; (b)IC ^k : in two combinations (A.AL A and B10.HTT B10.S[9R]) first grafts were rejected by day 30, although grafts to immunized mice showed a different pattern. In the third combination (B10.HTTB10.S[7R]) first grafts were retained but immunized mice rejected their grafts, (c)IC ^s : B10.S(9R) regularly reject B10.S(7R) first grafts, but immunized mice retain their grafts. In two other combinations first grafts were retained but grafts to immunized recipients were rejected; while in a third combination rejection did not occur at all. The background of the recipient appeared to be important in determining the variable pattern of rejection, and there is evidence for a similarity of the H-genes inIC ^s andIC ^k , and inIC ^k andIC ^p . Graft rejection occurred independently of known differences in Ia specificities, indicating thatH-2IC and the genes determining Ia specificities are probably different, although when grafts were performed in the presence of known la differences, graft rejection usually occurred.     

Evidence for a third,Ir-associated histocompatibility region in theH-2 complex of the mouse

Skin grafts transplanted from B10.HTT donors onto (A.TL × B10)F₁ recipients are rapidly rejected despite the fact that the B10.HTT and A.TL strains should be carrying the sameH-2 chromosomes and that both the donor and the recipient contain the B10 genome. The rejection is accompanied by a production of cytotoxic antibodies against antigens controlled by theIr region of theH-2 complex. These unexpected findings are interpreted as evidence for a third histocompatibility locus in theH-2 complex,H-2I, located in theIr region close toH-2K. The B10.HTT and A.TL strains are postulated to differ at this hypothetical locus, and the difference between the two strains is explained as resulting from a crossing over between theH-2 ^t1 andH-2 ^s chromosomes in the early history of the B10.HTT strain. TheH-2 genotypes of the B10.HTT and A.TL strains are assumed to beH-2K ^s Ir ^s / ^k Ss ^k H-2D ^d andH-2K ^s Ir ^k Ss ^k H-2D ^d , respectively. Thus, theH-2 chromosomes of the two strains differ only in a portion of theIr region, including theH-2I locus. The B10.HTT(H-2 ^tt) and B10.S(7R)(H-2 ^th) strains differ in a relatively minor histocompatibility locus, possibly residing in theTla region outside of theH-2 complex.     

Kidney transplantation between congenic versus standard inbred strains of rats: I The significance ofH-1 and non-H-1 gene differences

Concepts of “antigenic strength” in organ transplantation have been evaluated in relation to orthotopic kidney allografts inH-1 congenic strains of rats. Untreated recipients reject fully allogeneic kidneys possessing singleH-1 differences as acutely as kidneys displaying multiple histocompatibility differences. Heterozygozity for H-1 specificities as well as for H-1 plus non-H-1 specificities (semiallogeneic kidneys) favors long term survival (autoenhancement), especially when the specific immune response genes of the recipient lead to reduced reactivity. In active enhancement, the transplantedH-1 congenic kidneys, devoid of additional weak antigens, retain prolonged functional integrity. Weak non-H-1 antigens substantially influence the successful establishment of specific enhancement in an adverse way, either as additive immunogens or as target sites for the effector arm of the rejection response.     

Genetic predisposition of donors affects the allograft outcome in kidney transplantation; polymorphisms of stromal-derived factor-1 and CXC receptor 4

Genetic interaction between donor and recipient may dictate the impending responses after transplantation. In this study, we evaluated the role of the genetic predispositions of stromal-derived factor-1 (SDF1) [rs1801157 (G>A)] and CXC receptor 4 (CXCR4) [rs2228014 (C>T)] on renal allograft outcomes. A total of 335 pairs of recipients and donors were enrolled. Biopsy-proven acute rejection (BPAR) and long-term graft survival were traced. Despite similar allele frequencies between donors and recipients, minor allele of SDF1 rs1801157 (GA+AA) from donor, not from recipients, has a protective effect on the development of BPAR compared to wild type donor (GG) (P = 0.005). Adjustment for multiple covariates did not affect this result (odds ratio 0.39, 95% C.I 0.20–0.76, P = 0.006). CXCR4 rs2228014 polymorphisms from donor or recipient did not affect the incidence of acute rejection. SDF1 was differentially expressed in renal tubular epithelium with acute rejection according to genetic variations of donor rs1801157 showing higher expressions in the grafts from GG donors. Contrary to the development of BPAR, the presence of minor allele rs1801157 A, especially homozygocity, predisposed poor graft survival (P = 0.001). This association was significant after adjusting for several risk factors (hazard ratio 3.01; 95% C.I = 1.19–7.60; P = 0.020). The allelic variation of recipients, however, was not associated with graft loss. A donor-derived genetic polymorphism of SDF1 has influenced the graft outcome. Thus, the genetic predisposition of donor should be carefully considered in transplantation.     

Marrow allograft success inW/W v anemic mice: Relation to skin graft survival times

Genetically anemicW/W ^v mice were cured by marrow allografts from donors of 13 out of 18 tested strains that differed at non-H-2 histocompatibility alleles defined by skin or tumor grafting. They were also cured by donors from all four tested congenic lines whose antigenic differences had been defined by induction of serum antibodies. They were not cured acrossH-2 differences. Tail skin graft survival times on uncuredW/W ^v recipients were determined for all congenic lines used as marrow donors. The longest and shortest skin graft survival times predicted correctly marrow graft success or failure. NoW/W ^v mice were cured by marrow grafts from donors of the three congenic lines whose skin grafts were rejected in fewer than three weeks. Almost everyW/W ^v mouse grafted was cured by marrow grafts from donors of the 13 congenic lines whose skin grafts survived longest, from 11 to more than 25 weeks. Intermediate skin graft survival times failed to predict whether marrow grafts would succeed.W/W ^v mice were cured by marrow from four congenic lines with mean skin graft survival times of 4.2, 4.4, 8, and 9 weeks, while marrow grafts failed from other congenic lines with mean skin graft survival times of 3.3, 3.4, 4.8, and 8.7 weeks. The simplest explanation for these results is that the antigens specified by theH-2, H-3, H-4, H-25, andH-28 loci are strongly immunogenic on both marrow precursor cells and skin,H-17 andH-24 are strongly immunogenic on skin but not on marrow, andH-12 is strongly immunogenic on marrow precursor cells but less strongly on skin.     

The role of H-2 and non-H-2 antigens and genes in the rejection of murine cardiac allografts

Genes of the major histocompatibility complex (MHC) in the mouse (H-2 complex) have been shown to be an important factor in determining the immune responsiveness of various strains of mice to isolated antigens (e. g., lysozyme). The role of MHC genes in controlling the responsiveness of mice to multiple alloantigens is less well-defined, and although non-MHC genes have been shown to be important in determining responsiveness in some systems (e. g., haptens), they have not been demonstrated as yet to influence the rejection of vascularized organ allografts. In this study, the responsiveness of mice to vascularized cardiac allografts transplanted across well-defined major (H-2) and minor (non-H-2) histocompatibility barriers was investigated using congenic mice in 32 different donor/recipient combinations. The results show that both H-2 and non-H-2 gene products can act as target alloantigens for rejection. At the responder level, they may interact to effect responsiveness of a recipient strain to multiple alloantigens. In no case in this study has any one gene or group of genes been found to confer universal high or low responder status.     

Transfer of Infectious Drug Resistance in Microbially Defined Mice

Germ-free mice were intentionally associated with drug-resistant donor strains of Escherichia coli known to carry R factors and with drug-sensitive recipient strains. In vivo transfer of R factors was observed in all experiments, involving five different donor-recipient combinations. The number of converted recipients varied, depending upon the donor-recipient combination, but in all cases it was restricted by limiting numbers of either recipient or donor strains in the digestive tract of the microbially defined mice. Converted recipients were detected in fecal material as early as 5.5 hr after mice were associated with donor and recipient bacteria. Donors, recipients, and converted recipients were detectable in the stomach, small intestine, cecum, and large intestine of the microbially defined mice and their suckling young.     

Adoptive transfer of delayed-type hypersensitivity to Sendai virus. I. Induction of two different subsets of T lymphocytes which differ in H-2 restriction as well as in the Lyt phenotype

This paper investigates kinetics and antigenic and genetic requirements for transfer of delayed-type hypersensitivity (DTH) reactions against Sendai virus. Kinetics of sensitization of effector cells are similar to those described in other virus systems. Maximum DTH response is elicited in the footpad 7 days after sensitization; maximal increase in footpad thickness is found approximately 24 hr after injection of virus and transfer of immune lymphocytes. DTH effector cells are Ig^? and are susceptible to treatment with anti-theta antibody and complement. After transfer of immune lymphocytes a DTH reaction can be elicited by infectious virus, uv light-inactivated virus or cell-cultured Sendai virus which lacks fusion capacity. Requirements of H-2 compatibility are dependent on the biological nature of the virus preparation used for challenge: High doses of infectious or uv light-inactivated Sendai virus require K, D, or IA region compatibility; low concentrations of infectious virus require K and/or D region compatibility, while cell-cultured fusion-negative Sendai virus requires IA region homology. At the level of induction K, D region-restricted DTH-effector T lymphocytes (Td) and cytolytic T lymphocytes (Tc) are stimulated to a greater extent by infectious virus than by uv light-inactivated virus. Furthermore, stimulation of these T lymphocytes is dependent on the route chosen for immunization: intravenous (iv) injection is superior to intraperitoneal (ip) injection; subcutaneous (sc) injection causes the lowest degree of sensitization. IA region-restricted Td lymphocytes are stimulated to comparable amounts by infectious or uv light-inactivated Sendai virus independent of the route of immunization. Td lymphocytes, which require IA region compatibility and Td lymphocytes which require K or D region compatibility can be distinguished by their Lyt phenotype, e.g., IA region-restricted Td lymphocytes are characterized by Lyt-1⁺2^? antigens, while K or D region-restricted Td lymphocytes are phenotypically Lyt-1(⁺)2⁺.     

The specificity and significance of the inhibition of Fc receptor binding by anti H-2 sera

The possibility that Ia antigens are unique among H-2 antigens in their relationship to the Fc receptor was investigated in an EA rosette assay. Antibody specific for antigens in various regions of theH-2 complex was incubated with mouse cells, and the ability of the cells to form rosettes with antibody-coated chicken erythrocytes was tested. Antibody raised against the H-2 antigens of Ia-negative tumor cells was highly effective in inhibiting rosette formation. A variety of antisera againstK-, I-, andD-region antigens tested in recombinant mice inhibited EA rosette formation, suggesting that antigens in each of these regions could be detected in rosette inhibition. The F(ab′)₂ fragments of all antisera tested also produced specific EA rosette inhibition. Finally, antibody against Ia antigens failed to inhibit bone marrow RFCs, although antibody against H-2K and H-2D antigens did inhibit. Although H-2 serology is in a state of rapid change at present, it must be concluded that in this assay, antibody against antigens in theK andD regions as well as theI region can inhibit EA rosette formation. Inhibition of these rosettes by anti H-2 sera is therefore not due to a special association of Ia antigens with Fc receptors.     

Genetic resistance to murine leukemia induced by different radiation leukemia virus variants; a comparative study on the role of theH-2 complex

Association of resistance to leukemogenesis with theH-2 complex was investigated. Three variants of the radiation leukemia virus: D-RadLV, RadLV and RS-RadLV were tested. Using congenic and recombinant mouse strains, the region includingH-2K andI-A/B was found to be involved with resistance to all three viruses. In addition,H-2D-region linked resistance was found in leukemogenesis in two of the three passages. The phenotypic expression of resistance associated withH-2D seemed to depend on the simultaneous presence ofH-2K-H-21-linked resistance alleles. TheH-2 haplotypes associated with resistance or sensitivity to the different RadLV variants were different for each passage, suggesting that there is a large degree of heterogeneity both inH-2-linked resistance and between the radiation-induced leukemia viruses.     

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.     

The genetic control of the immune response to murine histocompatibility antigens

The genetic control of the immune response to H-4 histocompatibility alloantigens is described. The rejection of H-4.2-incompatible skin grafts is regulated by anH-2-linkedIr gene. Fast responsiveness is determined by a dominant allele at theIrH-4.2 locus. TheH-2 ^b ,H-2 ^d , andH-2 ^s haplotypes share the fast response allele;H-2 ^a has the slow response allele. Through the use of intra-H-2 recombinants, we have mapped theIrH-4.2 locus to theI-B subregion of theH-2 complex; theH-2 ^h4 ,H-2 ¹⁵, andH-2 ^t4 haplotypes are fast responder haplotypes. These observations suggest that the strength of non-H-2 histocompatibility antigens is ultimately determined by the antigen-specific recipient responsiveness.     

Study ofH-2 mutations in mice VI. M523, a newK end mutant

A newH-2 mutation was found in a mouse belonging to CBA/CaLacSto (H-2 ^k ) strain and designated 523, the proposed haplotype symbol for which isH-2 ^ka . The line CBA.M523 carries this mutation and is fully congenic with the parental strain, except for the mutant site 523. The mutation 523 is located within theK- end of theH-2 gene complex. Phenotypically, it causes prompt skin graft rejection and pronounced graft-versus-host activity in strain combination CBA/Sto⇄C-BA.M523. Attempts to produce active alloantisera in the same strain combination have so far been unsuccessful.     

Immune response to calf collagen type I in mice: A combined control ofIr-1A and nonH-2 linked genes

The genetically controlled immune response to calf skin collagen type I in mice could be demonstrated to be governed by at least two genes. One is linked to theH-2 complex and located within theIA subregion. High-responder alleles areH-2 ^b ,H-2 ^f , andH-2 ^s . The other gene(s) is not linked to theH-2 complex and high-responder allele(s) are found in the genome of B10 mice but not in the genome of DBA mice. There are strong indications that theIr-1A gene controls the response at the T-cell level, whereas it is assumed that the background gene(s) control the immune response at a different level.