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C S David

Publications and source records attributed to C S David.

At least 163 records · Page 9Linked to original sources
PubMed1990

Susceptibility of HLA-B27 transgenic mice to Yersinia enterocolitica infection.

The majority of patients with reactive arthritis have the major histocompatibility complex class I gene HLA-B27. The development of arthritis in these patients often occurs following infection with one of several enteric bacteria, including Yersinia enterocolitica. In this study, transgenic mice expressing HLA-B27 and their negative full sibs were infected intravenously with Yersinia enterocolitica 0:8 WA in an attempt to develop an experimental model of reactive arthritis. To date, no reactive arthritis has been observed; however, a significantly higher incidence of paralysis was observed in the HLA-B27+ transgenic mice. Injection of 10(5) organisms induced hind limb paralysis in 8 out of 30 of the HLA-B27 transgenic mice (27%) and in only 1 of the 24 negative siblings (4%). Paralysis occurred in 14 out of 30 HLA-B27+ mice (47%) at a dose of 10(4) organisms. Only 2 of the 25 negative siblings (8%) were affected at this dose. Paraspinal abscesses were found in all of the paralyzed animals. At the 10(4) dose most of the HLA-B27+ mice (70%) succumbed to the disease within 4 weeks, while the mortality in their B27- full sibs was less than 10%. Thus, HLA-B27 transgenic mice have higher mortality and morbidity from infection with Y. enterocolitica 0:8 WA than corresponding HLA-B27- littermates.

Animals↗
PubMed1990

Hemopoietic histocompatibility (Hh-1) regulatory and structural genes of the f haplotype map to H-2.

Natural resistance to bone marrow stem cell (BMC) grafts in lethally irradiated mice is a consequence of natural killer cell recognition and elimination of BMC that express hemopoietic histocompatibility (Hh) antigens inherited noncodominantly. The major Hh genetic region, Hh-1, maps to H-2. The phenotype of the BMC was determined by grafting BMC into panels of irradiated mice, and measuring splenic incorporation of the radioactive specific DNA precursor, 5-iodo-2'-deoxyuridine [125I] (IUdR) 5 days after cell transfer. Our previous analysis of Hh-1 antigen expression on BMC of intra-H-2 recombinant inbred strain mice indicated that Hh-1 regulatory genes (Hh-1r) map in the H-2S/H-2D interval. Nine recombinants described here shared the following: (1) the DL region was donated by H-2f that was associated with expression of determinant 2, shared with H-2d/Hh-1d, and (2) had crossovers in the S/D or Ea/S intervals or within Eb, and (3) had BMC that were Hh-1 null. F1 hybrids of crosses between mice of these recombinant strains with H-2d strain mice had BMC that expressed determinant 2, suggesting that a structural gene had been lost by the crossover event. Crosses with H-2b strain mice produced mice whose BMC were Hh-1 null, indicating that the H-2f/Hh-1f regulatory genes were still intact. We suggest that Hh-1r maps to the S/D interval of all haplotypes so far studied, even H-2f. The structural gene for the f haplotype is centromeric of Eb and may be Kf, based on recent data supporting the role of class I antigens in Hh antigen expression.

Animals↗
PubMed1990

Inhibition of alloantigen presentation by cyclosporine.

Cyclosporine (CsA) was examined for its ability to inhibit alloantigen presentation by spleen cells in a primary mixed lymphocyte reaction; by gamma interferon-induced P388.D1 macrophages to an alloreactive T cell clone; and by a B cell lymphoma, B1D.beta to an alloreactive T cell hybridoma. Alloantigen-presenting cells were treated with CsA or its inactive analogs for 2 hr, washed extensively (four times), and added to the T cells. Using this protocol, CsA maximally inhibited allorecognition by the T cells at 1000 ng/ml in all three systems. An HPLC assay for CsA cell failed to detect any significant CsA carryover into the T cell assays. Supernatant transfer experiments also failed to demonstrate CsA carryover in the more sensitive T cell hybridoma assay. These transfer experiments also failed to demonstrate the generation of inhibitory factors during the assay. Northern blot analysis and a cell-surface ELISA failed to observe any decreases in MHC class II induction in/on P388.D1 cells with CsA present during the induction. Due to the lack of detectable (less than 10 ng/ml) CsA carryover, we hypothesize that CsA has a direct effect on the formation of stimulatory MHC class II in our alloreactive systems.

Animals↗
PubMed1990

Genes for MHC, TCR and MIs determine susceptibility to collagen induced arthritis.

Type II collagen induced arthritis (CIA) in mice is an animal model for polyarthritis. The susceptibility to the disease is linked to the class II genes of H-2 gene complex (MHC). The susceptibility requires T cells expressing certain receptors coded by the V beta genes. Further, the MIs gene products in combination with the class II molecules can up- or down-regulate the T cells involved in the disease. The disease is mediated by the presentation of an arthritogenic epitope on the collagen type II peptide by the MHC class II molecule, which is recognized by a T cell expressing certain V beta receptors, triggering the autoimmune response. These studies point out possible mechanisms of rheumatoid arthritis in humans and suggest various methods of immune intervention to down-regulate the disease.

Amino Acid Sequence↗
PubMed1990

Molecular analysis of two recombinant mouse strains with crossovers in the E alpha recombination hot spot.

Recombination in the E alpha gene of the mouse major histocompatibility complex has been found to occur only in mice derived from crosses between strains with the k and p haplotypes. In the present paper the crossover sites of six additional strains were examined by RFLP analysis. These recombinant strains were derived from crosses between strains (B10.RPD1, B10.RPD2) with crossovers in the E alpha gene and B10.M (H-2f). Four have crossover sites between the E alpha gene and the S region. One recombinant (B10.RFD7) also crossed over within the E alpha gene and a second (B10.RPF1) crossed over within a segment located to the left of the E alpha gene. This study suggests that the DNA sequence responsible for recombination in the E alpha gene is also present in the B10.RPD1 and B10.RPD2 recombinant strains.

Animals↗
PubMed1990

Immunogenetic aspects of human thyroglobulin-reactive T cell lines and hybridomas.

The in vitro proliferative response to T cells primed with human thyroglobulin (Tg) was compared in 11 independent haplotypes on B10 background. B10.K and B10.S mice were the most responsive, whereas, with the exception of B10.PL (H-2u), all other B10 congenics were intermediate responders. The two best responders to in vitro challenge with human Tg, of the k and s haplotype, are the same as those showing H-2-linked susceptibility to induction of experimental autoimmune thyroiditis (EAT) with mouse Tg. Since shared epitopes on human and mouse Tgs have been shown to be thyroiditogenic by adoptive transfer studies in CBA (H2k) mice, the findings indicate that shared epitopes may be studied in appropriate (i.e. EAT-susceptible) strains of mice. Therefore, we proceeded to develop methods to produce T-cell lines and hybridomas to human Tg in B10.K and B10.S mice, test their cross-reactivity to heterologous Tgs and their Ia restriction patterns. By using antigen-presenting cells from recombinant strains, we identified restriction elements encoded by the I-A subregion alone and a combinatorial molecule from the I-A/I-E subregions.

Animals↗
PubMed1990

Major histocompatibility complex-conferred resistance to Theiler's virus-induced demyelinating disease is inherited as a dominant trait in B10 congenic mice.

Intracerebral inoculation of Theiler's murine encephalomyelitis virus into susceptible strains of mice produces chronic demyelinating disease in the central nervous system characterized by persistent viral infection. Immunogenetic data suggest that genes from both major histocompatibility complex (MHC) and non-MHC loci are important in determining susceptibility or resistance to demyelination. The role of the MHC in determining resistance or susceptibility to disease can be interpreted either as the presence of antigen-presenting molecules that confer resistance to viral infection or as the ability of MHC products to contribute to pathogenesis by acting as viral receptors or by mediating immune attack against virally infected cells. These alternatives can be distinguished by determining whether the contribution of the MHC to resistance is inherited as a recessive or dominant trait. Congenic mice with different MHC haplotypes on identical B10 backgrounds were crossed and quantitatively analyzed for demyelination, infectious virus, and local virus antigen production. F1 hybrid progeny derived from resistant B10 (H-2b), B10.D2 (H-2d), or B10.K (H-2k) and susceptible B10.R111 (H-2r), B10.M (H-2f), or B10.BR (H-2k) parental mice exhibited no or minimal demyelination, indicating that on a B10 background, resistance is inherited as a dominant trait. Although infectious virus, as measured by viral plaque assay, was cleared inefficiently from the central nervous systems of resistant F1 hybrid progeny mice, we found a direct correlation between local viral antigen production and demyelination. These data are consistent with our hypothesis that the immunological basis for resistance is determined by efficient presentation of the viral antigen to the immune system, resulting in local virus clearance and absence of subsequent demyelination.

Animals↗
PubMed1990

Effect of H-2 genes on expression of HLA-B27 and Yersinia-induced arthritis.

HLA-B*2705 transgenic mice were continuously backcrossed to mice of the B10 background with various haplotypes. A high level of expression of the HLA-B27 protein was detected on peripheral blood lymphocytes (PBLs) from mice homozygous for H-2b, H-2f, H-2s, H-2p, H-2r, and H-2k haplotypes by FACS analysis with the ME-1 antibody. A lower level of expression of B27 was observed on PBLs from H-2v mice. Little or no expression of B27 was detected on PBLs from H-2 or H-2d mice. We hypothesize that the HLA-B27 heavy chain is analogous to and competes with endogenous class I heavy chains in the H-2d, H-2q and H-2v haplotypes. Interestingly, other studies in our laboratory have demonstrated that mice with the H-2d and H-2q haplotypes with deletions of certain T cell receptor subsets are more prone to Yersinia-induced arthritis (YIA). Therefore, the mouse model of YIA may provide insights into the mechanisms of HLA-B27-linked spondyloarthropathies in man.

Animals↗
PubMed1990

Role of enterobacteria and HLA-B27 in spondyloarthropathies: studies with transgenic mice.

Several MHC (major histocompatibility complex) genes are associated with increased incidence of disease. The strongest association is between the class I gene, HLA-B27, and ankylosing spondylitis (AS). HLA-B27 is also highly associated with Reiter's syndrome. As not all subjects with HLA-B27 develop AS or Reiter's syndrome, environmental factors may have a key role in the pathogenesis of these arthritic diseases. Several studies have implicated klebsiella in the development of AS, whereas Reiter's syndrome may result from infection with yersinia, shigella, salmonella, campylobacter, or chlamydia. Transgenic mice present a unique opportunity to study the association of specific MHC genes and disease. HLA-B27 transgenic mice were produced to study the association of HLA-B27, bacterial infection, and arthritic disease. Mice with the HLA-B27 gene are more susceptible to intravenous infection with Yersinia enterocolitica 0:8 WA than negative litter mates as shown by a higher incidence of spinal abscesses and mortality. Understanding the mechanism(s) responsible for this difference may yield valuable insights into the pathogenesis of HLA-B27 associated disease in humans.

Animals↗
PubMed1989

MHC class II A alpha and E alpha molecules determine the clonal deletion of V beta 6+ T cells. Studies with recombinant and transgenic mice.

Interactions between MHC class II genes and minor lymphocyte stimulating (Mls) associated products are responsible for clonally deleting self-reactive T cells in mice. Here we demonstrate the role of the intact I-A and I-E molecules as well as the individual A alpha and E alpha chains in the deletion of cells bearing the V beta 6 TCR. DBA/1 (H-2q, Mls-1a) mice were crossed with various inbred congenic, recombinant, and transgenic strains and the F1's were screened for V beta 6 expression. All I-E+ strains were fully permissive in deleting V beta 6+ T cells. I-E- strains expressing I-A b,f,s,k,p permitted only partial deletion, while I-Aq strains showed no deletion. Recombinant I-Aq and I-Af strains which expressed E kappa alpha chain in the absence of E beta chain showed a decrease in V beta 6+ T cells as compared to their H-2q and H-2f counterparts. Furthermore, transgenic mice expressing E kappa alpha Aq beta gene in an H-2q haplotype (E kappa alpha Aq beta?) gave similar results to that of the recombinants in deleting V beta 6 T-cells. The role of the 1-A molecule was also shown by the partial deletion of V beta 6+ T cells in H-2q mice expressing transgenic I-Ak molecules. These results demonstrate that the E alpha chain is important in the deletion of V beta 6 T-cells in Mls-1a mice. The role of A alpha chain is also implied by the permissiveness of E kappa alpha Aq beta but not Aq alpha Aq beta molecules in the deletion of V beta 6+ T cells.

Animals↗
PubMed1989

In vivo expression and function of hybrid Ia dimers (E alpha A beta) in recombinant and transgenic mice.

We have found cell surface expression of an E alpha molecule in recombinant and transgenic mouse strains lacking an E beta molecule. Flow cytometry has shown low level expression of E alpha in B10.RQB3 (I-AqEk alpha) and B10.RFB2 (I-AfEk alpha) mice. We have also found that B10.Q (H-2q) mice can express the Ek alpha transgene. Since these strains do not have functional E beta chains, we propose that the E alpha A beta hybrid dimers are formed in low numbers and can be picked up by FACS analysis. So far we have not been able to identify these hybrid molecules by cytotoxicity or immunoprecipitation. The E alpha/A beta molecule can function in vivo during thymic selection in the clonal deletion of two V beta TCR subsets, V beta 11 and V beta 6, which have been shown to interact with the intact I-E molecule.

Animals↗
PubMed1989

RIII S/J (H-2r). An inbred mouse strain with a massive deletion of T cell receptor V beta genes.

We have identified an inbred strain of mouse, RIII S/J (H-2r), that has the largest known deletion of the TCR V beta genes by screening with mAb and TCR V beta specific probes. Upon screening of PBL with mAb F23.1, which is specific for V beta 8 TCR, RIII S/J was found to be negative. On further screening with mAb KJ 23a, which is specific for V beta 17a TCR, RIII S/J was completely negative. We next tested RIII S/J with mAb 44-22-1, which is specific for V beta 6 TCR, and found it also to be negative. The (B10 X RIII)F1 mice showed a 50% expression of V beta 6 gene, indicating a genomic rather than a clonal deletion. mAb KJ25, detecting V beta 3, was positive in RIII S/J, denoting the downstream boundary for the deletion. Southern blot analysis of liver DNA using TCR V beta-specific probes confirmed the deletion of V beta 8 gene subfamily and V beta 5 gene subfamily, along with V beta 9, V beta 11, V beta 12, and V beta 13 genes similar to the known TCR V beta deletion mutants (SWR, SJL, C57L, and C57Br). In addition, RIII S/J is missing V beta 6, V beta 15, and V beta 17 genes. Our mapping of the deletion indicates that RIII S/J has lost approximately 130 kb of V beta chromosome and with it 13 V beta genes out of the known 21 V beta genes of the TCR. The deletion is marked by the presence of V beta 10 gene upstream and V beta 3 gene downstream.

Animals↗
PubMed1989

Molecular analysis and repair of a defective Ef alpha gene.

The molecular basis for the defective expression of Ef alpha was determined by analysis of the 5' region of a full length Ef alpha gene. The gene was isolated from a genomic library prepared from the A.CA/SnDv mouse strain. DNA sequence analysis of the 5' portion of the Ef alpha gene, which encodes the 5' regulatory sequences and the signal peptide, revealed the presence of a stop codon in the exon encoding the signal peptide. The remainder of the sequences were highly related to sequences found in previously characterized, functional E alpha alleles. Previous studies indicate that the f allele is transcribed at rates comparable to the rates of functional alleles and that mRNA accumulates in the cytoplasm. Primer extension analysis demonstrated that Ef alpha transcripts initiate identically to the functional Ek alpha allele, mapping the defect in the Ef alpha gene 3' of the transcriptional initiation site. To determine whether the stop codon in the signal peptide was the only major defect in this gene, reciprocal chimeric genes were constructed in which the 5' regions, including the first exons, of the defective f allele and the functional k allele were exchanged. The hybrid genes were inserted into an SV40 promotor driven expression vector for cotransfection with an Ek beta gene. Surface I-E expression was demonstrated using I-E specific mAb in Cos-7 and L cell lines transfected with the hybrid gene consisting of the 5' region of the k allele and the 3' portion of the f allele. Therefore, the single stop codon present in the exon encoding the leader peptide of the Ef alpha gene appears to be the only defect preventing this gene from expressing a functional E alpha-chain.

Amino Acid Sequence↗
PubMed1989

Influence of complement C5 and V beta T cell receptor mutations on susceptibility to collagen-induced arthritis in mice.

SWR/J mice are resistant to collagen-induced arthritis (CIA) despite having a susceptible H-2q haplotype. We have earlier demonstrated the possible role of the V beta TCR mutation of SWR in the resistance to CIA. To investigate the influence of the C5 deficiency of SWR in this resistance, crosses were made between SWR and A/J (C5 deficient, TCRwild, H-2a), and between SWR and C3H.A (C5 sufficient, TCRwild, H-2a). Upon immunization with bovine type II collagen in adjuvant, there was a similar incidence and severity of arthritis in H-2q-bearing mice in the back-crosses A x (SWR x A) ad C3H.A x (SWR x C3H.A). The absence of hemolytic complement was confirmed in the arthritic A x (SWR x A) back-cross mice by standard SRBC hemolytic assays. In addition C57L (H-2b) mice, which were C5 sufficient but had a V beta TCR deletion mutation similar to SWR, could not complement for CIA susceptibility in H-2q-bearing C57L x (SWR x C57L) back-crosses and in (C57L x SWR)F2 hybrids. These studies show that complement C5 does not play a significant role in CIA susceptibility, and further implicate the V beta TCR mutation in the resistance to CIA in SWR mice.

Animals↗