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K L McCoy

Publications and source records attributed to K L McCoy.

At least 37 records · Page 2Linked to original sources
PubMed1995

delta 9-Tetrahydrocannabinol modulates antigen processing by macrophages.

delta 9-Tetrahydrocannabinol (THC) exposure suppresses multiple immunological functions of macrophages. The ability of macrophages exposed to THC to process and present soluble protein antigens was investigated by the stimulation of antigen-specific helper T cell hybridomas to secrete interleukin-2. The T cell response to hen egg lysozyme was dramatically reduced after a 24-hr pretreatment of a macrophage hybridoma with THC. In contrast, THC exposure did not alter the capacity of the macrophage hybridoma to process chicken ovalbumin and augmented their presenting cell function for a pigeon cytochrome c response. These findings could not be attributed to differential effects of THC on either cell viability or expression of the antigen receptor-associated CD3 complex by the T cells. The level of T cell activation with peptides of lysozyme and cytochrome c, which do not require processing, was inhibited only at the highest concentrations of THC, suggesting that THC mainly affects antigen processing. Peritoneal macrophages exposed to THC during an antigen pulse and fixed with paraformaldehyde showed similar effects on the subsequent T cell responses to lysozyme and cytochrome c in the absence of THC, arguing against a possible influence of THC on the T cells. Therefore, THC differentially modulates the capacity of macrophages to process antigens that is necessary for the activation of CD4+ T cells.

Animals↗
PubMed1993

Differences among various lineages of antigen-presenting cells in processing exogenous antigen internalized through transferrin receptors.

The Ag, pigeon cytochrome c, was coupled to human ferric transferrin by a heteroligation technique to target Ag into the endosomal transport pathway via transferrin receptors. The ability of various types of APC that do or do not express transferrin receptors to process exogenous Ag in their endosomes was investigated by the stimulation of Ag-specific CD4+ T cells with the transferrin-Ag conjugate in a serum-free assay. When two B lymphoma cells were the source of APC, the conjugate was significantly more potent than native Ag in activating the T cells, agreeing with our previous finding using a third B lymphoma cell. The conjugate and Ag were similarly presented by splenic B cells that lack transferrin receptors to the T cells. However, both a macrophage hybridoma and a MHC class II-L cell transfectant hardly elicited a T cell response to the conjugate, although a response to native Ag was readily observed. These findings could not be attributed to an absence of transferrin receptors or receptor-mediated internalization of the conjugate, nor to differential expression of MHC class II molecules or li chain by the APC. The poor presentation of the conjugate by the L cell transfectants was associated with diminished catabolism of the conjugate, however, the macrophage hybridoma rapidly degraded the conjugate, similar to the B lymphoma cell. Peritoneal macrophages, which lack transferrin receptors, and the macrophage hybridoma induced a response to the conjugate only at concentrations that allowed internalization by fluid phase pinocytosis. The lower potency of the conjugate compared with native Ag with non-B-presenting cells suggest that these cell types process the conjugate by a different mechanism than used by B cells. Differences in the mechanism of Ag processing used by APC of distinct cell lineages may possibly influence immune responsiveness.

Animals↗
PubMed1993

Antigen presentation by B lymphoma cells. Requirements for processing of exogenous antigen internalized through transferrin receptors.

The Ag, pigeon cytochrome c, was delivered to the early endosomes in a form coupled to human ferric transferrin that entered APC through transferrin receptors. The processing of the transferrin-Ag conjugate by B lymphoma cells was compared with that of unconjugated native Ag that entered APC by a nonreceptor-mediated mechanism. Within 5 min after internalization, catabolized conjugate was detected in isolated early endosomes and did not accumulate in these organelles. Analysis of the rapid catabolism of the conjugate demonstrated that the Ag, not the transferrin, portion of the molecule was degraded by the APC, suggesting that similar proteases may mediate the processing of the conjugate and native Ag. The processing mechanisms of these molecules shared similarities. Treatment of APC with chloroquine or paraformaldehyde interfered with the stimulation of Ag-specific CD4+ T cells by both transferrin-Ag conjugate and native Ag. However, the T cell responses to the conjugate and native Ag were different in two important respects. First, T cell activation by the conjugate began at an earlier time point and occurred at a faster rate than T cell stimulation by the same concentration of native Ag during a 3-h time course. Second, the T cell response to the conjugate, but not to native Ag, was diminished by treating APC with cycloheximide, a reversible protein synthesis inhibitor. This partial inhibition of the conjugate response by cycloheximide could not be attributed to significant effects on transferrin receptor expression, or on internalization, recycling, or degradation of the conjugate. The differential cycloheximide-sensitivity of the T cell responses indicates that the processing pathways of the two molecules are different. Our findings suggest that the early endosomes may function as an Ag-processing compartment, and that more than one pathway may lead to productive processing in B lymphoma cells.

Antigen Presentation↗
PubMed1993

Exogenous antigens internalized through transferrin receptors activate CD4+ T cells.

The role of endosomes in exogenous Ag processing was investigated by targeting Ag into the endosomal transport pathway via transferrin receptors. The Ag, pigeon cytochrome c and chicken OVA, were coupled to human ferric transferrin by a heteroligation technique. The conjugates were significantly more efficient than native Ag in stimulating Ag-specific CD4+ T cells, when the APC expressed transferrin receptors. The addition of ferric transferrin eliminated the enhanced response. Paraformaldehyde-fixed APC did not present the conjugates, indicating that the conjugates still required processing to activate T cells. An augmented level of T cell activation was not observed when the APC lacked transferrin receptors or when the conjugate contained the apoenzyme form of transferrin, which does not bind the receptor. The conjugate followed an intracellular pathway similar to that for transferrin, remaining in low density vesicles. Degraded conjugate appeared rapidly in culture supernatants, within 5 min, and peaked by 20 min; under these conditions a T cell response to the conjugate was elicited that was consistent with an early processing compartment. Our results suggest that antigenic peptide fragments can be generated in the early endosomes, without delivery of these Ag to the lysosomes. Thus, various Ag may have differential processing requirements, dictated by their molecular nature, that determine the site of Ag processing.

Animals↗
PubMed1991

Suppression of splenic accessory cell function in mice exposed to gallium arsenide.

Acute exposure of mice to a single intratracheal dose of gallium arsenide (50, 100, and 200 mg/kg) depresses the primary IgM antibody response to the T-dependent antigen sheep red blood cells (SRBC) through alterations in the function of splenic accessory cells. To determine the mechanism by which GaAs exposure influences splenic accessory cells, the cells were isolated by adherence and their functional capability investigated 24 hr following GaAs exposure in the animal. Splenic adherent cells from GaAs-exposed mice were greatly impaired in their ability to process and present the particulate antigen SRBC to a SRBC-primed T-cell population. However, GaAs exposure did not inhibit phagocytosis of fluorescent covaspheres by these cells, nor did it inhibit in vivo phagocytosis of 51Cr-labeled SRBC, indicating that the findings reported here were not due to decreased uptake of antigen by the accessory cells. Furthermore, production of IL-1 by these cells from exposed mice was not different from control and addition of exogenous IL-1 to cultures did not reverse GaAs-induced inhibition of the primary antibody response. GaAs exposure did not affect the percentage of Ia positive macrophages (F4/80 positive cells), but the amount of cell surface IAk molecules expressed was significantly decreased as measured by flow cytometry. In contrast to the SRBC response, GaAs did not suppress the ability of adherent splenocytes to process and present the antigen pigeon cytochrome c to the helper/inducer T cell clone F1.A.2 or the antigen KLH (keyhole limpet hemocyanin) to KLH-primed T cells. Therefore, GaAs exposure interferes with the capacity of splenic macrophages to process and/or present the particulate antigen SRBC, but not the soluble protein antigens pigeon cytochrome c or KLH.

Animals↗
PubMed1990

Contribution of endosomal acidification to antigen processing.

The role of endosomes in antigen processing was investigated with mutant cells, expressing a temperature-sensitive defect in endosomal acidification, as a source of antigen-presenting cells for CD4+ T cell responses. When the mutants were incubated at the nonpermissive temperature, their ability to process a variety of native antigens was greatly impaired. These cells could present peptide fragments normally, and the higher temperature did not affect antigen processing by the wild-type parental cells. The mutant cells do not have a defect in the internalization and overall degradation of native antigen. Upon return to the permissive temperature, the mutant cells rapidly recover their ability to process antigen. Therefore, acidified early endosomes have an important function in antigen processing, and other acidified organelles may also be involved.

Animals↗
PubMed1989

Diminished antigen processing by endosomal acidification mutant antigen-presenting cells.

The role of acidified endosomes in Ag processing was investigated using mutant Chinese hamster ovary cells that express temperature sensitive defects in their acidification mechanism. These cells were transfected with MHC class II genes to convert them to APC. When such mutant cells were incubated at the nonpermissive temperature, losing early endosomal but not lysosomal acidification, their ability to process several native protein Ag was impaired. The nonpermissive temperature did not affect Ag processing by transfected wild type parental cells. Furthermore, T cells were stimulated normally under these conditions when the mutant cells presented antigenic peptide fragments, which do not require processing. The mutant cells were also not defective in the uptake and overall degradation of native antigen. The elimination of Ag processing by paraformaldehyde and chloroquine treatment of the Chinese hamster ovary cells indicates that these cells do not use a different process than physiologic APC. These results suggest that acidification of early endosomes is an important event in Ag processing.

Animals↗
PubMed1988

The role of intracellular acidification in antigen processing.

The experiments described here were designed to investigate the role of intracellular acidic organelles in the processing of foreign antigens. Simultaneous addition of antigen and the inactive precursor of an acid protease, cathepsin L, greatly diminished the stimulation of a T-cell hybridoma specific for pigeon cytochrome c. The effect occurred during antigen processing as the presentation of a peptide fragment was unaffected by precursor-cathepsin L. The effect required uptake of the enzyme via mannose-6-PO4 receptors and was specific for antigenic determinants cleavable by the enzyme. The results confirm, without any pharmacologic manipulation, that foreign antigen moves into an acidic environment during some phase of its processing. We also explored the role in antigen processing of one acidic compartment, the early endosome, by examining CHO cells expressing a temperature-sensitive defect in endosomal acidification. These cells were transfected with MHC class II genes to convert them into APC for CD4+T cells. When these cells were incubated at the nonpermissive temperature, their ability to process antigen was impaired. There was no negative effect on presentation of antigenic peptides nor on the uptake and degradation of the antigen. Wildtype CHO cells similarly transfected and tested did not show any aberrancy in antigen processing. The inhibition of processing by the mutant cells was only partial, even under stringent conditions, and the residual processing was eliminated by chloroquine. Our findings provide the first evidence for a contribution by acidified early endosomes to the pathway of antigen processing and suggest that other acidified compartments are also involved.

Acids↗
PubMed1986

Tolerance defects in New Zealand Black and New Zealand Black X New Zealand White F1 mice.

The susceptibility of autoimmune NZB and (NZB X NZW)F1 mice to the induction of tolerance by monomeric BSA was compared with several normal mouse strains. Unresponsiveness in T and B lymphocyte compartments was probed by challenging with DNP8BSA and measuring anti-DNP and anti-BSA antibodies separately. Tolerance induced by monomeric BSA was carrier specific, and there was no evidence of epitope-specific suppression. Normal NZW, NFS, and B10.D2 mice were easily rendered tolerant with monomeric BSA and did not produce anti-DNP or anti-BSA antibodies after challenge with DNP8BSA. By contrast, the lack of anti-DNP antibody response in similarly treated NZB mice was dependent on the dose of monomeric BSA, indicating that the helper T cells were partially resistant to tolerance induction. NZB mice treated with a high dose of monomeric BSA produced anti-BSA, but not anti-DNP, antibodies after immunization. Thus, the anti-carrier B cells in NZB mice may have been primed by monomeric BSA. The presence of the xid gene on the NZB background rendered the mice susceptible to induction of tolerance, suggesting that the tolerance defect in NZB mice involves the B cell compartment. This abnormal antibody response was a dominant trait: (NZB X NFS)F1 and (NZB X B10.D2)F1 mice had the same characteristics as NZB mice. These F1 hybrids do not develop autoimmune disease, indicating that resistance to experimental tolerance induction expressed at a B cell level may not be sufficient for disease development. In contrast to NZB and other NZB F1 hybrids, (NZB X NZW)F1 hybrids treated with monomeric BSA and challenged with DNP8BSA responded to both DNP and BSA. The contribution of a B cell defect to the tolerance abnormality of (NZB X NZW)F1 mice was examined by analyzing the effect of the xid gene on the progeny of (NZB.xid X NZW)F1 mice. Unlike the effect of the xid gene on NZB mice, both phenotypically normal heterozygous female and phenotypically xid hemizygous male mice produced anti-DNP and anti-BSA antibodies after tolerance induction and immunization, demonstrating that a major helper T cell abnormality was present in (NZB X NZW)F1 mice. The (NZW X B10.D2)F1 hybrid was rendered tolerant by this procedure, indicating that the helper T cell defect (NZB X NZW)F1 mice may have resulted from gene complementation with the NZB mice contributing partial resistance of T helper cells to tolerance induction.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1986

Increased amplifier T cell activity in autoimmune NZB mice and its possible significance in the expression of autoimmune disease.

The expression of amplifier- and helper-T cell activity was examined in NZB/N mice of different ages. Amplifier T cell activity develops in a cyclic manner; it decreases between 16 and 35 wk of age and then increases to maximal levels in mice older than 50 wk of age. The loss of suppressor T cell activity and increased amplifier T cell activity coincide with the development of autoimmune diseases in aging NZB/N mice. Although helper T cell activity is evident in young NZB/N mice, it is absent in old NZB/N mice expressing maximal amplifier T cell activity. This provides additional support for the fact that amplifier and helper functions are mediated by different subpopulations of T cells.

Age Factors↗
PubMed1985

Effects of the motheaten gene on murine B-cell production.

The rapidly fatal autoimmune disease in the mutant mouse known as motheaten is caused by an autosomal recessive gene and is characterized by hypergammaglobulinemia and autoantibody production, among other defects. The cellular kinetics of B-cell maturation were investigated in three-week-old motheaten mice and their normal littermates to determine whether any abnormality in cell production of the B lineage could be correlated with B-cell hyperactivity. The production rates and renewal times of newly produced bone marrow, splenic small B-lymphocytes, and splenic plasma cells were examined by in vivo tritiated-thymidine administration using a pulse-chase protocol and radioautography of immunofluorescence-stained cells. Because small B-lymphocytes in both organs were produced at comparable rates in the mutant mice and in their normal littermates, primary B-cell production was unaffected in the mutant mice. In contrast, splenic plasma cells were produced 10-30 times faster in motheaten mice than in normal mice. The enhanced rate of plasma cell production in motheaten mice could be correlated with a concurrent increased loss of labeled large B-lymphocytes, presumably rapidly dividing activated B cells. Thus, the excessive antibody production in motheaten mice may be reflected by the increased plasma cell production.

Animals↗
PubMed1985

Two defects in old New Zealand Black mice are involved in the loss of low-dose paralysis to type III pneumococcal polysaccharide.

Treatment of normal mice with a subimmunogenic dose of type III pneumococcal polysaccharide (SSS-III) results in the development of an antigen-specific state of unresponsiveness termed low-dose paralysis. This unresponsiveness is mediated by T suppressor cells and can be transferred by Lyt-2+ T cells, but not by L3T4+ T cells, obtained 18 hr after priming. As autoimmune New Zealand Black (NZB) mice age, there is a progressive decrease in low-dose paralysis to SSS-III. The defect in older NZB mice resulting in decreased suppressive activity was investigated by transferring primed Lyt-2+ T cells from young into old mice, and vice versa. Enlarged Lyt-2+ T cells from old NZB mice could not suppress the SSS-III response of young recipients. However, Lyt-2+ T cells of normal cell size were efficient in inhibiting the antibody response upon transfer. Primed Lyt-2+ T cells from young NZB mice did not affect the response of old recipients, but effectively suppressed the response of young mice. These results suggest that there are two defects involved in the decline of low-dose paralysis to SSS-III in aging NZB mice: Enlarged Lyt-2+ T cells may lose their ability to function as mediators of suppression; and B cells may become resistant to T cell-mediated suppression.

Aging↗
PubMed1985

Enlargement of Lyt-2-positive T cells is associated with functional impairment and autoimmune hemolytic anemia in New Zealand Black mice.

We investigated the relationship between the increased cell diameter of Lyt-2+ T cells and the development of autoimmune disease in aging NZB and NZB X NZW F1 hybrid (BW) mice. Individual animals were analyzed for Lyt-2+ T cell size (by narrow-angle forward light scatter), anti-erythrocyte autoantibodies, anemia, proteinuria, and splenomegaly. The peak light scatter of the Lyt-2+ T cells correlated with the level of anti-erythrocyte autoantibodies and severity of hemolytic anemia, but not with proteinuria or splenomegaly. The cell size of this T cell subset did not increase in old BW or in NZB mice homozygous for the xid gene (NZB.xid). The in vivo administration of bacterial lipopolysaccharide to young NZB mice did not stimulate the enlargement of Lyt-2+ T cells. Ly-2+ T cells from old NZB mice could be stimulated by concanavalin A (Con A) to express interleukin 2 (IL 2) receptors and to synthesize DNA in vitro. However, in vivo administration of Con A to old NZB mice did not induce the expression of IL 2 receptors on Lyt-2+ T cells. Further, in vivo T suppressor function was impaired in old NZB mice with enlarged Lyt-2+ T cells. Thus, the enlargement of Lyt-2+ T cells in old NZB mice appears related to impaired T cell function in vivo and is associated with the development of anti-erythrocyte autoantibodies and autoimmune hemolytic anemia.

Aging↗
PubMed1984

Spontaneous production of colony-stimulating activity by splenic Mac-1 antigen-positive cells from autoimmune motheaten mice.

Cultured splenocytes from 3-wk-old autoimmune motheaten mice (me/me) spontaneously produced colony-stimulating activity (CSA), which stimulated the formation of bone marrow myeloid colonies. The production of CSA was not dependent on the presence of serum; this activity was not produced by spleen cells from their phenotypically normal littermates (+/-) or from other normal mouse strains. The peak level of CSA occurred early during the culture period, and within 48 hr the activity was markedly diminished. Cell fractionation studies demonstrated that cells expressing Mac-1 antigen produce CSA and are most likely to be mononuclear phagocytes. The unusual proliferative capacity in vitro of splenic mononuclear phagocytes from motheaten mice probably results from the spontaneous production of CSA by Mac-1 antigen-positive cells. Defective regulation of the production of monokines may contribute to the severity of the immunologic disease of these mutant mice.

Animals↗
PubMed1983

Accelerated rate of mononuclear phagocyte production in vitro by splenocytes from autoimmune motheaten mice.

The rapidly fatal autoimmune disease accompanied by pneumonitis in the mutant mouse known as motheaten is caused by an autosomal recessive gene. The proliferative capacity and production rate of splenic mononuclear phagocytes at different maturational stages, defined by morphologic criteria, were examined by two in vitro tritiated thymidine administration protocols and radioautography. The replicative rate of splenic promonocytes from 3-week-old normal mice was found to approximate that of adult bone marrow cells. Monocytes, macrophages, and previously described macrophagelike cells from motheaten mice had an accelerated rate of maturation in vitro, compared with similar cells from normal mice. These differences in the production rates could be attributed to the shorter period of time that promonocytes from the mutant mice are in the S phase of the cell cycle. Evidence, also, verified that the macrophagelike cells were derived from promonocytes via the monocyte-macrophage compartment. The unusual in vitro growth characteristics of the splenic mononuclear phagocytes from motheaten mice could be a manifestation of an in vivo abnormality of this lineage that could contribute to the development of pulmonary disease in these animals.

Animals↗
PubMed1982

Abnormal in vitro proliferation of splenic mononuclear phagocytes from autoimmune motheaten mice.

Motheaten mice develop combined immunodeficiency and fatal autoimmune disease that follow autosomal recessive inheritance. In splenocyte cultures of motheaten mice, supplemented with 5% normal serum proliferating cells (MP) were present exhibiting morphologic characteristics of mononuclear phagocytes at light and electron microscopic levels. The macrophage nature of these cells was confirmed by the lack of Thy-1 antigen and immunoglobulins; the expression of Mac-1 antigen, FcR for IgG, and Ia antigens on their cell surfaces; their ability to phagocytize EA and adhere to plastic; the presence of nonspecific esterase and lysomal enzymes in their cytoplasm; and the pattern of peroxidase localization similar to monocyte-derived macrophages. MP from motheaten mice exponentially grew in culture in the absence of exogenous growth factors with a doubling time of approximately 76 hr. Although these cells were present in splenocyte cultures of normal controls, their number did not increase during the culture period under the same conditions. The addition of dextran sulfate further enhanced the proliferation of MP from motheaten mice, and induced exponential growth of these cells from normal controls, reaching only the level of unstimulated cells from motheaten mice. Radioautographic analysis demonstrated that MP substantially contributed to the elevated spontaneous and dextran sulfate-induced DNA synthesis in splenocyte cultures. Therefore, the in vitro abnormality of MP may be indicative of in vivo aberrancies of macrophages from motheaten mice and lends credence for investigating the role of macrophages in immunodeficiency and autoimmunity that develop very early in motheaten mice.

Animals↗