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Biomedical subjects

D K Bishop

Publications and source records attributed to D K Bishop.

At least 55 records · Page 3Linked to original sources
PubMed1996

Direct transfer of a foreign MHC gene into human melanoma alters T cell receptor V beta usage by tumor-infiltrating lymphocytes.

The direct introduction of foreign genes into tumors shows promise as a therapeutic modality to enhance tumor immunogenicity. Hence, melanoma nodules were directly injected with a vector encoding an allogeneic MHC class I molecule, HLA-B7. Tumor-infiltrating lymphocytes (TIL) were isolated from cutaneous melanoma biopsies before and after HLA-B7 gene transfer. TIL were expanded in interleukin-2 (IL-2) by standard techniques for approximately 4 weeks, then analyzed for T cell receptor V beta usage by quantitative reverse transcriptase polymerase chain reaction (RT-PCR). Prior to gene transfer. TIL V beta usage was found to be highly restricted, the only one to four V beta families being expressed and one or two of these families representing more than 90% of the repertoire. As anticipated, TIL V beta usage varied among patients expressing different HLA types. However, V beta 13 was over-represented in that six of eight patients utilized V beta 13 as a dominant family, regardless of HLA type. Following HLA-B7 gene transfer, TIL V beta usage was markedly altered: (1) V beta families that dominated following gene transfer differed from the V beta families utilized by TIL prior to treatment, and (2) introduction of the HLA-B7 gene resulted in a more diverse repertoire with an increase in the number of V beta families represented. In two patients, TIL were evaluated before treatment and from multiple, distinct melanoma nodules following gene transfer. In these two patients, a comparison was made between TIL V beta profiles obtained after treatment from nodules that had been injected with the HLA-B7 gene or left untreated. Interestingly, the V beta repertoires of TIL from uninjected nodules following gene transfer were similar to that of TIL from injected nodules, rather than pretreatment TIL. These data demonstrate a direct immunological effect of foreign MHC gene transfer into human melanoma, and suggest that local expression of an allogeneic MHC molecule generates systemic alterations in the antitumor immune response.

CD4 Antigens↗
PubMed1996

Ovarian response after gonadotropin treatment of heifers immunized against gonadotropin-releasing hormone.

Angus x Hereford heifers were used to determine whether the ovaries of heifers immunized against GnRH would respond to exogenous gonadotropins. Heifers (n = 23; 24 mo; 491 +/- 23 kg) were assigned to a 2 x 2 factorial experiment: immunization against GnRH conjugated to human serum albumin (GnRH-IMM) or human serum albumin alone (HSA-IMM), and treatment with gonadotropins (GTH) or saline. The GTH treatment was 2,000 IU PMSG + 1,000 iu hCG, and control heifers were given saline (s.c.). Immunization against GnRH (GnRH-IMM) caused cessation of ovulation (progesterone < 1 ng/mL plasma for 3 wk) and GTH treatment began within 3 to 5 wk after heifers became anovulatory. Heifers immunized against HSA (HSA-IMM) were treated between d 9 and 11 of the estrous cycle. All heifers were given PGF (i.m.) on d 2 (d 0 = start of GTH treatment) and 2,000 IU of hCG (i.m.) on d 4. Ovaries of heifers were evaluated by ultrasound. Blood was sampled daily from d 0 to d 8 and every second day through d 24 to quantify progesterone and estradiol. Concentrations of estradiol increased with time after GTH but the response was reduced by GnRH-IMM (IMM x GTH x day, P < .01). Number of follicles > or = 6 mm (d 0 to 14) was increased after GTH (GTH x day, P < .001) but was decreased (P < .03) by GnRH-IMM. Immunization against GnRH reduced total follicles > or = 6 mm on d 4 in heifers on both saline and GTH treatments. Treatment with gonadotropins increased the number of follicles > or = 16 mm on d 14 compared with control heifers. The number of CL was increased by GTH (GTH x day, P < .001) but the response was reduced (P < .03) in GnRH-IMM heifers compared with HSA-IMM heifers. We conclude that treatment with gonadotropins will induce follicular growth and ovulation in heifers that are anovulatory due to immunization against GnRH.

Animals↗
PubMed1996

T cell receptor (TCR) repertoire in alloimmune responses.

A large number of alloantigenic determinants could be generated by both the direct and indirect alloantigen presentation pathways. Hence, a heterogeneous population of T cells expressing a wide variety of receptors would be expected to respond to this diverse array of alloantigenic determinants. However, T cells expressing highly restricted T cell receptor (TCR) variable genes have been reported in a variety of alloimmune responses. A similar phenomenon has been observed in a wide variety of other immune responses, from those induced by superantigens, to very specific responses induced by a single peptide presented by a single MHC molecule. Given this scenario, the limited number of T cell clones which dominate an allograft rejection response, or for that matter an autoimmune response or a tumor specific response, could be therapeutically targeted by virtue of the selected TCR expression.

Antigen Presentation↗
PubMed1995

In vivo depletion of CD8+ T cells results in Th2 cytokine production and alternate mechanisms of allograft rejection.

A current hypothesis states that Th1 cytokines promote allograft rejection and that Th2 cytokines promote graft acceptance. We present data that question the tolerogenic activity of Th2 cytokines, and we suggest that Th2 cytokines may evoke allograft rejection by recruitment of alternate effector mechanisms. Unmodified rejection of mouse heterotopic cardiac allografts is associated with the accumulation of large numbers of donor-reactive CD8+ CTL within the allograft, which is indicative of a Th1-driven cellular response. However, when recipients are depleted of CD8+ CTL, rejection still occurs and is associated with an aggressive cellular infiltrate rich in eosinophils, large mononuclear cells, and fibroblast-like cells. Eosinophils, which are responsive to the Th2 cytokines IL-4 and IL-5, are not present in unmodified rejecting allografts. Differential production of Th1 versus Th2 cytokines was further suggested by altered levels of IgG2a (promoted by IFN gamma) and IgG1 (promoted by IL-4) alloantibody in the sera of these mice; IgG2a dominated the alloantibody response in unmodified allograft recipients, whereas IgG1 levels increased in recipients depleted of CD8+ CTL. Altered intragraft cytokine gene expression was verified by RT-PCR; Th1 (IL-2, IFN gamma), but not Th2 (IL-4, IL-5, IL-10), cytokine mRNAs were readily detectable in the allografts of unmodified recipients. In contrast, both Th1 and Th2 cytokine genes were expressed in the allografts of mice depleted of CD8+ CTL. These data suggest that donor-reactive CD8+ CTL inhibit intragraft production of Th2 cytokines, thereby promoting a Th1 dominated-rejection response. Elimination of CD8+ cells allows Th2 cytokine production, which may have deleterious, rather than protective, effects.

Animals↗
PubMed1995

Evidence that human cardiac allograft acceptance is associated with a decrease in donor-reactive helper T lymphocytes.

We have reported that acute cardiac allograft rejection is associated with increased numbers of donor-reactive helper T lymphocytes (HTL) in the peripheral blood of patients. Further, increased frequencies of circulating donor-reactive HTL may predict allograft rejection episodes diagnosed by endomyocardial biopsy. The present study evaluates the relationship between donor-reactive HTL and allograft "acceptance" in cardiac transplant recipients bearing long-term allografts (> 1 year). Patients were categorized as either long-term acceptors or persistent rejecters based on the number of rejection episodes and the ability to withdraw steroid therapy. Limiting dilution analysis for IL-2-producing HTL was utilized, with cadaver donor splenocytes as a source of donor alloantigens. Donor-reactive HTL frequencies were determined from peripheral blood samples obtained before transplant, and at 1 month and 1 year after transplant. Individuals who accommodated their allografts and were withdrawn from steroid therapy had reduced numbers of donor-reactive HTL at 1 year after transplant as compared with earlier time points. Further, PBMC obtained from these individuals at 1 year after transplant responded weakly to donor alloantigens in a mixed lymphocyte response (MLR). This relationship between donor-reactive HTL and allograft accommodation was exemplified in a cardiac/liver transplant patient who was diagnosed with progressive multifocal leukoencephalopathy and removed from all immunosuppression. No subsequent rejection episodes were diagnosed. Donor-reactive HTL were not detectable and this individual failed to mount an MLR to donor alloantigens. However, a vigorous donor-reactive response was observed when MLR cultures were supplemented with exogenous IL-2. Therefore, nonresponsiveness to the allograft appeared to be due to a deficit in IL-2 production. In contrast, patients who experienced persistent rejection episodes and required continued steroid therapy maintained large numbers of donor-reactive HTL at 1 year after transplant. PBMC from these individuals responded vigorously to donor alloantigens in an MLR. Hence, monitoring donor-reactive HTL may identify individuals who have accommodated their graft and may tolerate a reduction in immunosuppression.

Graft Rejection↗
PubMed1995

Dissociation of mouse cardiac transplant rejection and donor alloantigen-specific T cell responsiveness.

Mouse hearts transplanted into MHC disparate donors are usually rejected 1 week after placement. It is widely accepted that alloantigen-reactive helper T lymphocytes (HTL) and cytotoxic T lymphocytes (CTL) are the key mediators of acute allograft rejection. This report demonstrates that the presence or absence of 'traditional' graft-reactive HTL and CTL is not necessarily related to allograft survival. In these studies, donor/recipient combinations disparate only at MHC or only at minor histocompatibility (mH) loci were employed. Allograft survival was monitored, donor-reactive IL-2 (interleukin-2) producing HTL and CTL were quantified by modified limiting dilution analysis, and serum levels of cytolytic alloantibody were determined. C57BL/10 hearts (H-2b) transplanted into B10.BR (H-2k) recipients (full MHC disparity) enjoyed prolonged survival despite massive infiltration of the allograft by donor-reactive HTL and CTL. IgM, but not IgG, donor-reactive alloantibody was present in the sera of these mice. Hence, traditional IL-2 producing HTL and CTL were not capable of mediating acute graft rejection, nor of providing help for alloantibody isotype switching in this MHC disparate combination. In contrast, C3H/HeN (H-2k) hearts transplanted into B10.BR (H-2k) recipients (mH disparity only) were acutely rejected. Donor-reactive HTL and CTL were rare or not detectable in these recipients, and cytolytic alloantibody was not detectable. Similar observations were made when B10.BR hearts were transplanted into C3H/HeN recipients. Interestingly, treatment of recipients with anti-CD4 monoclonal antibody prevented rejection of mH disparate allografts. Therefore, CD4+ T cells were required for rejection of mH disparate allografts, but this process was independent of detectable IL-2 production or CTL function. Hence, the significance of monitoring 'traditional' T cell functions should be questioned in certain donor/recipient combinations.

Animals↗
PubMed1995

Effects of growth rate on carcass composition and lipid partitioning at puberty and growth hormone, insulin-like growth factor I, insulin, and metabolites before puberty in beef heifers.

The effect of three rates of gain on carcass composition, lipid partitioning, age and BW at puberty, and concentrations of growth hormone (GH), IGF-I, insulin, glucose, and NEFA in plasma were evaluated in 38 Angus x Hereford heifers. Heifers were allotted by BW and age to three treatments with a replication in each of 2 yr: full-fed (n = 13; FF) to gain 1.36 kg/d; limit-fed (n = 12; LF) to gain .68 kg/d; maintenance-full-fed (n = 13; MFF) to gain .23 kg/d for 16 wk, then full-fed to gain 1.36 kg/d. Heifers were slaughtered within 10 d after the onset of puberty. At slaughter, kidney, pelvic, and heart fat (KPH) and udder (UDDER) were separated from carcass, as was fat surrounding viscera (OM). After 48 h at 4 degrees C a carcass side was dissected into subcutaneous fat (SC), intermuscular fat (SEAM), soft tissue (SFT = inseparable lean and fat), LEAN, and BONE. In yr 1, LF heifers (431 d) were older (P < .05) than MFF heifers (371 d) at puberty, but age of FF heifers (389 d) did not differ (P > .10) from that of LF and MFF heifers. In yr 2, FF heifers (351 d) were younger (P < .05) than LF and MFF heifers (398 and 434 d, respectively). The FF heifers had greater (P < .05) BW and a greater (P < .01) percentage of lipid in the carcass at puberty than LF and MFF heifers. During the first 16 wk of treatment, concentrations of NEFA were greater in heifers with slower daily gains (MFF > LF > FF; P < .01). Concentrations of NEFA were lesser and concentrations of IGF-I and insulin were greater in plasma of FF than in that of MFF heifers during the 10 wk before puberty. Treatment significantly altered age, BW, carcass composition, and lipid partitioning at puberty in beef heifers. We conclude that the percentage of body fat is not the sole regulator of puberty, and age may be an important modulator in determining the onset of puberty in beef heifers.

Aging↗
PubMed1994

RecA homologs Dmc1 and Rad51 interact to form multiple nuclear complexes prior to meiotic chromosome synapsis.

Dmc1 and Rad51, yeast homologs of the E. coli RecA protein, are shown by immunostaining to localize to as many as 64 sites within spread meiotic nuclei. Genetic requirements for this punctate pattern suggest it represents recombination intermediates. Dmc1 and Rad51 colocalize and are therefore likely to act together during recombination. Despite their similarities, the two proteins have specialized functions: Dmc1 complexes do not form in rad51 mutants, while Rad51 complexes are retained indefinitely in dmc1 mutants. Dmc1 and, by inference, Rad51 form complexes before synapsis as monitored by immunostaining for Zip1 protein. Analysis of zip1 mutants shows that Zip1 promotes dissociation of Dmc1 complexes. Colocalization of Dmc1 and Zip1 raises the possibility that Dmc1 and Rad51 are components of recombination nodules.

Cell Compartmentation↗
PubMed1994

Helper T lymphocyte unresponsiveness to cardiac allografts following transient depletion of CD4-positive cells. Implications for cellular and humoral responses.

Initial treatment of heterotopic cardiac transplant recipients with anti-CD4 mAb promotes long-term (> 60 days) allograft survival. We have used modified limiting dilution analysis to quantitate donor alloantigen-reactive helper T lymphocytes (HTL) and CTL in mice bearing long-term cardiac allografts. Despite repopulation of lymphoid tissues with CD4+ T cells, donor alloantigen-reactive IL-2 producing and IL-4-producing HTL were rare or not detectable in lymphoid tissues or in the graft. While donor-reactive precursor CTL were present in lymphoid tissues, modified limiting dilution analysis revealed that stimulated ("antigen-conditioned") CTL were not detectable, and few CTL were present in the graft. In addition, antibodies reactive with donor alloantigens were not detectable in the sera of mice bearing long-term cardiac allografts. To determine whether additional in vivo stimulation with donor alloantigens would elicit an immune response, sponge allografts were implanted into mice bearing long-term cardiac allografts. Previous reports from this laboratory have demonstrated that T cell infiltration of sponge allografts is dependent upon antigen-driven cytokine production. While third-party sponge allografts were readily infiltrated by third-party-reactive HTL and CTL, sponge allografts of the same strain as the cardiac allograft were not infiltrated by T cells. However, donor strain sponge allografts induced an IgM (but not IgG) alloantibody response. These data indicate that IgM alloantibody could be induced in the absence of a cellular response to the sponge allograft. Kinetic studies revealed that a transient IgM (but not IgG) response was induced by the initial cardiac transplantation in the absence of CD4+ cells. These IgM alloantibodies disappeared by day 21 despite the persistence of the allograft. These observations indicate that transient depletion of CD4+ T cells induces allograft-specific T cell tolerance, but does not eliminate the ability to mount an allograft-specific IgM response. To our knowledge, this is the first report of a transient humoral response to alloantigens that occurs in the absence of CD4+ T cells, and can be reinduced in "tolerant" animals that fail to mount a cellular immune response. Potential mechanisms involved in the development and maintenance of anti-CD4 mAb-induced tolerance are discussed.

Animals↗
PubMed1994

Body energy reserves influence the onset of luteal activity after early weaning of beef cows.

The influence of body energy reserves on the onset of luteal activity and concentrations of LH and IGF-I in serum was evaluated in postpartum anestrous beef cows after early weaning. Multiparous Hereford and Hereford x Angus cows (n = 24) were fed during gestation to establish body condition scores between 3 and 6 (BCS, 1 = emaciated; 9 = obese) at parturition. Concentrations of progesterone in plasma were determined weekly for 5 wk postpartum (PP). Anovulatory cows and their calves (n = 19) were confined in stalls on d 40 +/- 3 PP. Jugular cannulas were inserted on d 44 +/- 3 PP, and calves were weaned (d 0) the following day. Blood samples were collected from all cows for 4 h (every 10 min) before weaning and on d 1, 2, 4, 6, 8, and 10 after weaning and LH was quantified. Progesterone was quantified in daily blood samples until d 10, and in samples taken twice weekly until d 46. Within 25 d after weaning, 100% of the cows with BCS < or = 5 at weaning (n = 7) had initiated luteal activity, whereas only 43% (P < .01) of the cows with BCS < 5 (n = 12) had luteal activity. Mean serum IGF-I concentrations were correlated with BCS (r = .50; P < .05). Frequency of LH pulses was influenced (P < .01) by body condition at weaning but was not influenced by day after weaning. The number of LH pulses at weaning, serum IGF-I, and the interval to the onset of ovarian activity after early weaning of anestrous beef cows were influenced by BCS.

Anestrus↗
PubMed1993

Structural relationship of bacterial RecA proteins to recombination proteins from bacteriophage T4 and yeast.

RecA protein is essential in eubacteria for homologous recombination and promotes the homologous pairing and strand exchange of DNA molecules in vitro. Recombination proteins with weak sequence similarity to bacterial RecA proteins have been identified in bacteriophage T4, yeast, and other higher organisms. Analysis of the primary sequence relationships of DMC1 from Saccharomyces cerevisiae and UvsX of T4 relative to the three-dimensional structure of RecA from Escherichia coli suggests that both proteins are structural homologs of bacterial RecA proteins. This analysis argues that proteins in this group are members of a single family that diverged from a common ancestor that existed prior to the divergence of prokaryotes and eukaryotes.

Adenosine Triphosphatases↗
PubMed1993

CD4-positive helper T lymphocytes mediate mouse cardiac allograft rejection independent of donor alloantigen specific cytotoxic T lymphocytes.

Mouse heterotopic cardiac allograft recipients were depleted of CD4+ or CD8+ T lymphocytes in vivo to assess cellular requirements for graft infiltration, tissue damage, and acute allograft rejection. Modified limiting dilution analysis was employed to quantitate IL-2-producing Th lymphocytes (HTL) and CTL infiltrating the graft. Results were correlated with graft function and histologic evidence of tissue damage. In unmodified recipients, large numbers of donor alloantigen-specific CTL infiltrated the graft, overshadowing a modest number of HTL. CTL infiltration coincided with tissue damage and loss of graft function, suggesting a key role for CTL in rejection. In vivo treatment with anti-CD4 mAb inhibited both HTL and CTL infiltration, and no histologic evidence of tissue damage was observed. This observation suggested that HTL, although few in number, regulated the development of effector CTL and/or entry of these CTL into the graft. Reconstitution of HTL-depleted recipients with IL-2 resulted in graft infiltration by stimulated CTL, as assessed by modified limiting dilution analysis. However, these stimulated CTL failed to mediate tissue damage, and graft survival was prolonged. Unlike CTL obtained from unmodified recipients, graft-infiltrating CTL of IL-2-reconstituted mice were incapable of directly lysing donor cells in a 51Cr release assay. Hence, while IL-2 facilitated partial CTL differentiation and mobilization to the graft, additional signals appear necessary for maturation into lytic CTL. Furthermore, in recipients depleted of CTL by treatment with anti-CD8 mAb, HTL infiltrating the allograft, though few in number, were associated with extensive tissue damage and loss of graft function. These data suggest a less important role for CTL in the rejection process, and indicate that graft-infiltrating CTL are insufficient as sole mediators of cardiac allograft rejection. Potential mechanisms by which CD4+ HTL mediate cardiac allograft rejection independent of CTL are discussed.

Animals↗
PubMed1993

Pulsatile infusion of gonadotropin-releasing hormone initiates luteal activity in nutritionally anestrous beef cows.

Thirty-five nonlactating Hereford and Hereford x Angus cows weighing 469 +/- 8 kg, with a body condition score (BCS) of 5.5 +/- .6 and exhibiting normal estrous cycles, were fed to lose 1% of BW per week (n = 27) or to maintain BW (n = 8). Blood samples were collected every 7 d to monitor luteal activity and BW and BCS were recorded every 2 wk. When concentrations of progesterone in plasma were < 1 ng/mL for three consecutive weeks, anestrous cows (BCS, 3.1 +/- .7; BW loss, 24 +/- 5%) were assigned to pulses (i.v.) of saline (2 mL every hour), GnRH-4 (2 micrograms of GnRH every 4 h), or GnRH-1 (2 micrograms of GnRH every hour). Cows were confined to stalls for 4 d and two jugular cannulas were inserted 2 d before infusion of GnRH. Infusions began on d 0 and continued through d 14. Progesterone was quantified in daily blood samples and LH was quantified in samples collected every 10 min for 4 h on d-1, 0, 2, 4, 6, 8, 10, 12, 14, and 15. On d 16, cows were returned to pasture and blood samples were collected every 2nd d through d 30. Pulsatile infusion of GnRH increased (P < .05) the amplitude of LH pulses but did not affect pulse frequency. During infusion of GnRH every hour or every 4th h, concentrations of LH in serum were increased compared with those in saline-treated cows (P < .001), and cows receiving GnRH hourly had greater (P < .005) LH than cows given GnRH every 4 h.(ABSTRACT TRUNCATED AT 250 WORDS)

Anestrus↗
PubMed1992

DMC1: a meiosis-specific yeast homolog of E. coli recA required for recombination, synaptonemal complex formation, and cell cycle progression.

DMC1 is a new meiosis-specific yeast gene. Dmc1 protein is structurally similar to bacterial RecA proteins. dmc1 mutants are defective in reciprocal recombination, accumulate double-strand break (DSB) recombination intermediates, fail to form normal synaptonemal complex (SC), and arrest late in meiotic prophase. dmc1 phenotypes are consistent with a functional relationship between Dmc1 and RecA, and thus eukaryotic and prokaryotic mechanisms for homology recognition and strand exchange may be related. dmc1 phenotypes provide further evidence that recombination and SC formation are interrelated processes and are consistent with a requirement for DNA-DNA interactions during SC formation. dmc1 mutations confer prophase arrest. Additional evidence suggests that arrest occurs at a meiosis-specific cell cycle "checkpoint" in response to a primary defect in prophase chromosome metabolism. DMC1 is homologous to yeast's RAD51 gene, supporting the view that mitotic DSB repair has been recruited for use in meiotic chromosome metabolism.

Amino Acid Sequence↗
PubMed1992

Cyclosporin A and FK506 mediate differential effects on T cell activation in vivo.

Modified limiting dilution analysis techniques were used to evaluate the effects of the immunosuppressants cyclosporin A (CsA) and FK506 on alloantigen-induced T cell activation in vivo. Treatment of sponge matrix allograft recipients with either CsA or FK506 inhibited lymphocytic infiltration of the allograft, a process thought to be dependent on local lymphokine production. In addition, both immunosuppressants markedly reduced the absolute number of lymphocytes recovered from the draining lymph nodes (LN) and prevented CTL activation in the LN. However, Ag-primed helper T lymphocytes (HTL) were present in the draining LN of sponge allograft recipients treated with CsA, but not in recipients treated with FK506. T cell depletion experiments were performed to determine the phenotype of primed HTL in the LN of untreated and CsA-treated sponge allograft recipients. In untreated sponge allograft recipients, CD4+ and CD8+ Ag-primed HTL were present in the draining LN in equivalent numbers. In contrast, the majority of primed HTL in the LN of CsA-treated sponge allograft recipients were CD8+, rather than CD4+ T cells. These observations indicate that CsA and FK506 exert distinct in vivo effects at the level of HTL priming, and CD4+ and CD8+ HTL exhibit differential sensitivity to CsA in vivo.

Animals↗
PubMed1992

Rapid activation-independent shedding of leukocyte L-selectin induced by cross-linking of the surface antigen.

L-selectin (also termed LAM-1, Leu-8,TQ-1, gp90MEL-14, peripheral lymph node homing receptor and LECAM-1) is an adhesion protein thought to be important in leukocyte entry into lymphoid tissues and sites of inflammation. We, as well as others, have shown that leukocyte activation by chemotactic factors results in rapid shedding (release) of L-selectin from the cell surface. Here we have used flow cytometry, enzyme-linked immunosorbent assay, and SDS-PAGE/Western blot analysis to determine whether cross-linking of L-selectin in the absence of activation causes shedding. We found that rapid loss of leukocyte L-selectin expression (down-regulation) could be induced by treating cells with a chemical cross-linker [bis (sulfosuccinimidyl) suberate]. L-selectin down-regulation via cross-linking could occur at 4 degrees C and in the absence of detectable cell activation (increased expression of CD11b/18). The loss of L-selectin expression was due to shedding of the molecule from the leukocyte cell surface. Cross-linking of L-selectin with specific monoclonal antibodies also caused loss of surface expression of L-selectin at 37 degrees C. Finally, shed L-selectin was detected in the plasma of healthy adults whose peripheral blood leukocytes demonstrated no obvious signs of activation. Our results suggest that activation-independent shedding of leukocyte L-selectin may occur in vivo and a possible mechanism could involve cross-linking of leukocyte L-selectin. This provides a novel mechanism for rapid regulation of expression of a leukocyte-endothelial cell adhesion receptor.

Animals↗
PubMed1992

Mobilization of T lymphocytes following cardiac transplantation. Evidence that CD4-positive cells are required for cytotoxic T lymphocyte activation, inflammatory endothelial development, graft infiltration, and acute allograft rejection.

Modified limiting dilution analysis (LDA) techniques were used to evaluate the mobilization of antigen-stimulated helper T lymphocytes (HTL) and cytotoxic T lymphocytes (CTL) following allogeneic heterotopic cardiac transplantation. These modified LDA techniques allow a quantitative comparison of T cells that have been stimulated by antigen in vivo versus unstimulated precursor T cells of the same antigen specificity. Endothelial changes associated with mononuclear cell infiltration of the transplant were studied using endothelia-specific monoclonal antibodies and immunohistochemistry. Early (day 3) infiltration of cardiac allografts was characterized by a prevalence of donor alloantigen-specific HTL over CTL. Immunohistology revealed that the day-3 infiltrate was associated with areas of differentiated vascular endothelium, located primarily in the subepicardial region. Though donor-specific precursor HTL and CTL were present in the peripheral lymphoid tissues and blood, very few of them had been stimulated at this early time. During the latter phases of the response (days 6-9), antigen-stimulated HTL and CTL were present in the rejecting heart with CTL dominating the response. Accumulation of large numbers of donor-specific CTL in the allograft correlated with extensive inflammatory endothelial development, myocyte destruction, and loss of graft function by day 9. Stimulated HTL and CTL were detectable in peripheral lymphoid tissues at days 6 and 9. In addition, a marked increase in the number of donor-specific precursor CTL, but not precursor HTL, was observed in the lymphoid tissues at the peak of the response. Depletion of class II MHC-restricted T cells by in vivo treatment with anti-CD4 mAb eliminated HTL activity in all lymphoid compartments assessed and markedly reduced the number of CTL infiltrating the allograft. In addition, no stimulated CTL were detectable in lymphoid tissues, and the number of precursor CTL was not increased. In anti-CD4-treated recipients, cardiac allografts remained functional with minimal histological evidence of rejection for at least 21 days. Though graft-associated inflammatory endothelia were absent in anti-CD4-treated recipients at day 6, endothelial differentiation was observed in day 21 allografts in anti-CD4-treated recipients. These observations indicate that inflammatory endothelial development may precede T cell infiltration and subsequent loss of the cardiac allograft function. Thus, CD4-positive HTL are required for (1) graft-associated inflammatory endothelial development; (2) CTL activation in peripheral lymphoid tissues; (3) CTL accumulation in allografted tissues; and (4) acute cardiac allograft rejection.

Animals↗