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D V Cramer

Publications and source records attributed to D V Cramer.

At least 127 records · Page 7Linked to original sources

Genetic aspects of cellular interactions in the immune response.

Our understanding of the complex cellular interactions responsible for mediating effective immune responses has increased substantially in recent years. It is now clear that the genetic loci that control the interaction of the cells of the immune response encode groups of closely related cell-surface molecules. These molecules are the class I and class II antigens of the MHC, the differentiation antigens on lymphocyte subpopulations, and the receptors of various types, including the membrane immunoglobulin of B lymphocytes and the antigen receptors of T lymphocytes. Biochemical analysis of these cell surface molecules has demonstrated that they display important DNA sequence homologies. A polypeptide of approximately 110 amino acids comprises the basic building block for many of the cell surface molecules. Gradually, as a consequence of evolutionary development, the immune system has expanded its ability to respond to the external environment by an increased complexity of lymphocyte subpopulations and the surface structures that modulate their interaction. These cell surface molecules provide the structures that allow collaborative interaction of different cell types and that form the multiprotein receptor complexes involved in the recognition of, and specific response to, foreign antigens. Our future understanding of the control of the immune response will depend upon establishing the biochemical nature and the multifaceted interactions of these important molecules.

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Orientation of the loci encoding RT1.B polypeptides in the major histocompatibility complex of the rat.

The major histocompatibility complex of the rat (RT1) contains genes that code for two class II histocompatibility antigens. The r12 rat strain (WRC) was derived from a mating which revealed a genetic recombination that defines the two loci, RT1.B and RT1.D, that code for the class II antigens. The RT1.B and RT1.D antigens of the RT1 complex are protein dimers consisting of an alpha and beta glycoprotein chain and are homologous to I-A and I-E genes, respectively, in the H-2 complex of the mouse. We have performed Southern blot analysis on liver DNA from the r12 and parental strains to examine the precise location of the recombinant event. After digestion with restriction enzymes, the DNA was separated on agarose gels, blotted onto nitrocellulose, and hybridized with mouse H-2 cDNA probes specific for I-A alpha and beta genes. The pattern of restriction fragment polymorphisms demonstrated that the site of recombination is between the RT1.B alpha and the RT1.B beta genes. As a result of these findings, we suggest that the sequence of genes within the RT1 complex consists of RT1.A ... RT1.B beta ... RT1.B alpha ... RT1.D (alpha, beta) ... Other class I genes, possibly corresponding to mouse Qa and Tla-like genes, were also apparent in these experiments.

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Linkage of loci encoding a kidney endothelial antigen and fumarate hydratase (Fh-1) in the rat.

In the rat a single locus, provisionally designated Eag-1, controls the expression of an antigen present on the endothelium of kidney peritubular capillaries and veins. We have examined the linkage relationship between Eag-1 and 10 polymorphic loci including hemoglobin b, fumarate hydratase, peptidase-3, urinary pepsinogen, seminal vesicle protein, glycerophosphate dehydrogenase, esterase-1, esterase-6, pinkeye, and hooded. Tissue samples from animals derived from (AUG X BN.1C)F1 X AUG and (AUG X BN.1C)F1 X BN.1C backcrosses were examined and a linkage association between Eag-1 and Fh-1 (EC 4.2.2.1) was detected. The linkage distance between Eag-1 and Fh-1 is 21 cM (chi 2 = 27.9; p = 0.00001) and this association defines the third locus in the tenth (X) linkage group of the rat.

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The genetics of bone marrow transplantation in the rat.

We have used a variety of standard inbred, recombinant, and congenic rat strains to establish the effect of MHC and non-MHC genetic incompatibilities on bone marrow transplantation. The role of these loci in the successful establishment of bone marrow engraftment was first determined by examining the potential of donor marrow to protect recipient rats from a lethal dose of the myeloablative drug busulfan. Similar donor/recipient combinations were then used to examine the effects of the same incompatibilities on the induction of fatal graft-versus-host disease (GVHD) in recipients conditioned with a combination of busulfan and cyclophosphamide. The data obtained indicate that: (1) a difference for the entire MHC of the rat is sufficient to prevent marrow engraftment and to produce fatal GVHD; (2) the class I RT1.A locus of the rat MHC has a differential effect on bone marrow transplantation, since disparity for this locus prevents successful marrow engraftment, while this gene has little effect on the development of fatal GVHD; (3) disparity for the class I RT1.E locus has no effect on bone marrow engraftment and does not stimulate GVHD; (4) disparity for the class II locus, RT1.D, can prevent marrow engraftment and elicit fatal GVHD; and (5) incompatibility for non-MHC genes can prevent the establishment of bone marrow engraftment and elicit fatal GVHD.

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Chronic graft-versus-host disease in rats after syngeneic bone marrow transplantation.

A disease similar to the chronic graft-versus-host disease (cGVHD) seen following transplantation of human bone marrow was observed after syngeneic and allogeneic bone marrow transplantation in rats. Bone marrow grafts were exchanged between donors and recipients that were syngeneic or genetically different for the RT2 erythrocyte antigen locus by the use of AUG/AUG.2B and PVG/PVG.2A congenic pair donor/recipient strain combinations. After an initial period of well-being (120-180 days posttransplantation), several AUG and AUG.2B recipients of syngeneic or RT2-mismatched bone marrow developed clinical signs compatible with cGVHD. The clinical signs of the disease included: erythema, diffuse alopecia, thickened skin folds, and conjunctivitis. Laboratory findings included peripheral blood eosinophilia and impaired in-vitro proliferative responses to third-party spleen cells in the mixed lymphocyte reaction. Histological examination of the tissues of a limited number of rats with cGVHD showed subepidermal mononuclear inflammation with atrophy of the epidermis and adnexa of the skin, as well as plasmacytic hyperplasia of the lymphoid tissues. None of the PVG or PVG.2A recipients of syngeneic or RT2-mismatched marrow developed cGVHD. The development of cGVHD in AUG.2B recipients of syngeneic marrow and the absence of the disease in reciprocal marrow grafts between the PVG/PVG.2A rat strains suggests that the development of the disease in the AUG and AUG.2B recipients of RT2-mismatched bone marrow is not due to the RT2 disparity, but may be due to an autologous immune reaction. Furthermore, the finding that the cGVHD is only observed when the AUG and AUG.2B strains are used as recipients--not when the PVG or PVG.2A strains are used as recipients--suggests that the development of the disease is associated with the genetic background of the host and is independent of the background of the donor. It is possible that the use of high-dose cyclophosphamide treatment is involved in the pathogenesis of cGVHD, because the disease is observed only when the recipients are conditioned for transplantation with this immunosuppressive agent.

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Differences in DMBA-induced mammary neoplastic responses in two lines of Sprague-Dawley rats.

It has been reported that female Sprague-Dawley rats obtained from a U.S. source and studied in the U.S. gave a larger and more rapid mammary neoplastic response to radiation than did female Sprague-Dawley rats obtained from a Dutch source and studied in The Netherlands. To learn if the different mammary neoplastic responses of the two 'lines' of Sprague-Dawley rats are due to inherent differences between the lines of rats or due to differences in experimental conditions, two groups of rats from the American source and one group from the Dutch source were studied for their response to a chemical carcinogen, dimethylbenzanthracene (DMBA), at the same laboratory. When 10 mg of DMBA per 100 g body wt was given by stomach tube to 28 rats from the Dutch source, 367 days later approximately 25% of these rats had developed mammary carcinomas and approximately 18% had developed mammary fibroadenomas. When the same dose of DMBA was given to rats from the U.S. source, 300 days later 90 and 100% had developed mammary carcinomas and 83 and 95% had developed mammary fibroadenomas. Similar trends were found for the number of neoplasms per rat and the mean time of appearance of the neoplasms. It was concluded that there are inherent differences between Sprague-Dawley rats obtained in the U.S. and Sprague-Dawley rats obtained in The Netherlands in regard to their mammary neoplastic responses to DMBA, as well as in their responses to radiation. Genetic differences between the two lines were confirmed by establishing dissimilarities in the expression of erythrocyte antigens coded for by RT1 (major histocompatibility complex).

9,10-Dimethyl-1,2-benzanthracene↗

Genetic polymorphism of the sixth component (C6) of rat complement.

The complement protein C6 has been shown to be genetically polymorphic in the rat. Isoelectric focusing of plasma samples from 19 inbred strains demonstrated two electrophoretically distinguishable migration patterns, each consisting of three bands. Breeding studies with the use of the BN and DA strains showed that the C6 patterns were inherited in a manner consistent with the co-dominant autosomal expression of two alleles (C6 A and C6 B). The distribution of the C6 alleles in a backcross mating was compared with eight independently segregating marker genes: RT1.A, RT2, Gdc -1, Igk-1, Hbb, Svp-1, Fh-1, and Es-6. There was no detectable linkage between C6 and any of these eight loci.

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Linkage of quantitative variation of a serum protein (Sp) to the major histocompatibility complex of the rat.

An antiserum made in rabbits against a rat serum protein (provisionally designated Sp) has been used to establish the existence of a partially inbred line of animals that have low levels of this protein in their serum. This line of animals, the WRD line, was derived from a cross between a wild rat, WRF 6, and a rat of the F344 inbred strain. Offspring from this mating were selected for homozygosity at one wild major histocompatibility complex (MHC) haplotype and inbred for nine generations by brother-sister matings. When tested by immunodiffusion or rocket immunoelectrophoresis with a rabbit anti-rat Sp serum, the sera of these animals showed low levels of the protein when compared with other wild or inbred lines. Animals from the WRD line were backcrossed to the WF inbred strain and normal levels of the Sp protein were transmitted as an autosomal dominant trait. The gene controlling the serum levels of the protein is tightly linked to the MHC (RT1 complex) of the rat.

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Biochemical characterization of Ia antigens encoded by the RT1.B and RT1.D loci in the rat MHC.

Ia antigens in rats are genetically associated with typical MHC-linked immune response (Ir) genes. One rat Ir gene (IR-GLT) has recently been mapped to the RT1.D locus in a rat MHC recombinant, WRC. We have studied the expression of Ia antigenic determinants in WRC and its parental strains BN and WRA using a panel of monoclonal antibodies. Our results suggest that the inheritance of IR-GLT corresponds with the inheritance of I-E-like, but not I-A-like, antigens in the WRC rat. These observations were confirmed when WRC I-E-like antigens were analyzed by two-dimensional gel electrophoresis. We propose that RT1.D shares both functional and antigenic homologies with the mouse I-E subregion.

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A Class II monoclonal antibody specific for the RT1.B, rather than the RT1.D, product of the rat major histocompatibility complex.

An allospecific monoclonal antibody, 79.7.5., has been shown to be specific for a Class II histocompatibility product of the ACI (RT1.AaBaDa) rat. To further specify the reaction of this antibody to the B or D locus Class II products of RT1, we examined the binding of this antibody to peripheral blood lymphocytes (PBLs) of the WRC rat (haplotype RT1.AnBnDa). Radiolabelled monoclonal antibody 79.7.5. did not bind to PBLs from the WRC rat, but it did bind to PBLs from the WRA rat (RT1.AdBaDa) and the DA (RT1.AaBaDa) rat. These results were confirmed using radiolabelled Staphylococcus protein A in an indirect binding assay. In addition, binding of 79.7.5 could be inhibited by alloantiserum BN anti-BN.1A (DA) (directed against AaBaDa) but not by BN anti WRC (directed against Da). These data demonstrate that monoclonal antibody 79.7.5. reacts with the a allele product of RT1.B rather than RT1.D. This antibody can be used to probe the structures and functional roles of different Class II products of the rat major histocompatibility complex.

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The relative roles of MHC and non-MHC genes in heart and skin allograft survival.

Although the role of the major histocompatibility complex (MHC) of the rat (RT1) in graft rejection has been established, the role of non-RT1 genes is not well understood. To investigate the influence of MHC and non-MHC genes in graft rejection, various combinations of congenic and inbred strains of rats were used as donors and recipients of skin grafts and perfused abdominal heart grafts. In addition, hemagglutinating and cytotoxic antibody responses were evaluated to assess loci that were serologically active in transplantation. Our results demonstrate that: (1) RT1 antigens are the most important determinant in heart and skin rejection; (2) antigens controlled by non-MHC genes also play a major role in rejection because they cause disparate heart grafts to be rejected by day 18 and skin grafts by day 26; (3) RT2 cell antigens alone do not cause graft rejection; and (4) allogeneic differences at the RT1, RT2, RT3, and RT6 loci elicit an antibody response in heart transplantation.

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Comparison of antigens in the rat MHC that act as CML determinants.

We have examined a variety of RT1 haplotypes using congenic strains of rats to investigate the comparative relationships among MHC antigens that have been reported to serve as cell-mediated lympholysis (CML) targets. These studies demonstrate that: (1) RT1.A antigens can act as CML determinants, and in some haplotypes they display identical serological and CML reaction patterns; (2) in some donor-recipient strain combinations RT1.A antigens may act as immunodominant determinants, preventing the generation of effectors capable of recognizing other antigens, and (3) a portion of the antigenic determinants that define the anti-CT CML response can be mapped to the same region of the rat MHC as the RT1.E locus.

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CML characterization of a product of a second class I locus in the rat MHC.

In the rat, genes that control the expression of target antigens detected by cell-mediated lympholysis (CML) are present in the major histocompatibility complex (MHC). The relationship of these loci, CT and Ag-L, to each other and to other loci within the MHC is unknown. In this report, we demonstrate the existence of a CML target antigen in the (DA X BN)F1 anti-DA.1I(BI) strain combination. The gene coding for this antigen is linked to the RT1 complex as indicated by the CML reactivity of targets from backcross and congenic animals. Inhibition studies demonstrated that this antigen has the widespread tissue distribution characteristic of class I antigens, and the gene coding for this CML antigen maps coincident with the RT1.E class I locus as indicated by the lysis of targets from the recombinant strains r10 and r11. The CML can be blocked by antisera directed against a product of the RT1.E locus. The locus controlling this CML reactivity, like CT and Ag-L, has been separated from RT1.A by recombination; unlike CT and Ag-L, the produce of this CML locus appears to be identical with an RT1.E allelic product that has been serologically identified and biochemically characterized.

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Demonstration of a new genetic locus in the major histocompatibility system of the rat.

A new recombination within the major histocompatibility complex (RT1) of the rat has been detected. The recombination occurred between a wild-derived haplotype, provisionally designated p1, and the RT1 haplotype of the BN strain. The recombinant haplotype, designated p3, carries the RT1. A locus (classical histocompatibility antigens) of the BN strain, a locus from the BN strain that codes for the expression of an Ia antigen and strong mixed lymphocyte response (MLR), and a second locus derived from the p1 haplotype that controls the expression of a second Ia antigen, the ability to elicit a strong MLR and the immune response to poly(Glu52Lys33Tyr15). This recombinant therefore demonstrates the division of the RT1.B region into two loci, tentatively designated RT1.B and RT1.D, and provides evidence for the existence of at least four loci in the MHC of the rat.

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