Biomedical subjects

C R Stokes

Publications and source records attributed to C R Stokes.

At least 73 records · Page 4Linked to original sources

PubMed1992

Experimental proliferative glomerulonephritis in the cat.

A model of chronic serum sickness was used to induce immune-complex glomerulonephritis in seven experimental cats, by daily intravenous inoculation of an increasing dose (5 to 35 mg) of human serum albumin (HSA). At week four, two of the seven animals developed anterior uveitis. At week 23, two different animals developed the subcutaneous oedema characteristic of the nephrotic syndrome (NS), whilst the other five cats appeared clinically normal. The kidneys were examined at necropsy by light microscopy and by transmission electron microscopy. The glomeruli of four animals (three with both proteinuria and uraemia, and one with proteinuria only) showed morphological changes under light microscopy. The abnormalities suggested that a diffuse mesangial proliferative glomerulonephritis (GN) had been induced in three cats and diffuse membranoproliferative GN induced in another. Ultrastructural studies revealed electron-dense deposits (immune-complexes) in six of the seven cats. Two cats without glomerular abnormalities by light microscopy had mesangial deposits and three cats with mesangial proliferative GN had deposits at mesangial, subendothelial and/or subepithelial sites. The single cat with membranoproliferative GN had deposits at mesangial, subendothelial, subepithelial and intramembranous sites. Immunohistological examination (peroxidase-antiperoxidase technique) showed that HSA and immunoglobulin (IgG and IgM) were deposited in the glomeruli of these cats. Deposits were the most dense in cats with more severe renal lesions. Deposits of IgM were most abundant. An extensive cellular infiltrate, comprising macrophages, neutrophils and plasma cells, was observed only in the four animals which showed abnormalities in glomerular ultrastructure. The disease induced in these cats thus appears to differ from the membranous nephropathy previously described in the cat and bears a close resemblance to immune complex (IC) disease in man. In view of the relatively few specific animal models of IC-mediated proliferative GN, this model has potential for application to the study of human IC disease.

Animals↗

PubMed1992

Antibody response and antibody affinity maturation in cats with experimental proliferative immune complex glomerulonephritis.

An experimental model of proliferative glomerulonephritis (GN) in the cat, which closely resembles human proliferative forms of GN, has been used to study the role of antibody and antibody affinity in the development of immune complex-mediated renal disease. The serum IgG and IgM antibody response to antigen, average antibody affinity (avidity) and affinity heterogeneity of the IgG and IgM populations was assessed at varying times after commencement of chronic immunization with the antigen, human serum albumin (HSA), by enzyme immunoassay. Cats could be classified according to whether they were "low", "intermediate" or "high" IgG responders, by quantification of serum IgG values. Cats with the lowest serum IgG values failed to develop glomerulonephritis. However, there was no relationship between actual IgG values and the severity of the induced disease. In contrast to IgG, there was no division of cats into low or high IgM anti-HSA responders. Again, cats with the lowest IgM values failed to develop GN, but, more interestingly, a late, marked increase in serum IgM anti-HSA occurred only in cats that developed clinical signs of GN (anterior uveitis and nephrotic syndrome). Maturation of average, functional IgG affinity (avidity) for HSA following chronic immunization was clearly demonstrated for all cats. At the end of the experiment, all cats had IgG of high affinity for HSA and the average affinity heterogeneity of the IgG populations was less than in measurements taken earlier. Values of IgG affinity at the end of the experiment were very similar both in cats which developed GN and in those which remained clinically, biochemically and pathologically normal. In contrast to IgG antibody, some cats developed IgM of increased affinity, whilst others produced antibody of reduced affinity, following chronic immunization. There was no correlation between the development of disease and the production of either low or high affinity IgM antibody. Data indicated that an alteration in IgM affinity occurred at a late stage, as serum IgM levels increased, in cats which progressed to develop GN. These findings suggested that an increase in both serum IgG and IgM anti-HSA values, in particular IgM, was associated with the development of a more severe immune complex renal disease in these cats. Although there was no evidence that differences in the average affinity of either the IgG or IgM antibody populations for HSA, were associated with the development of disease, the increase in IgM values was also accompanied by a concomitant alteration in IgM affinity for antigen.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

PubMed1992

MHC class II expression in the bovine mammary gland.

The distribution of major histocompatibility complex (MHC) class II positive cells within the connective tissue and the epithelium of the involuted bovine mammary gland has been determined. The effect of intramammary administration of the antigens ovalbumin and formalin killed Streptococcus uberis on the distribution pattern has also been investigated. Infusion of formalin killed S. uberis increased cellular expression of class II antigens when compared with quarters either infused with ovalbumin, not infused at all, or from which minor pathogens had been isolated. The increased expression occurred particularly in the area of the gland cistern-secretory tissue junction.

Animals↗

PubMed1992

The intestinal and serum humoral immune response of mice to systemically and orally administered antigens in liposomes: I. The response to liposome-entrapped soluble proteins.

The development of oral vaccines is of great importance in veterinary medicine and new adjuvants and carriers are essential to this aim. Liposomes are effective systemic adjuvants but the relatively little data on their potential as oral adjuvants is inconclusive. Liposomes containing ovalbumin (OA) were effective adjuvants when administered intraperitoneally to mice. Feeding mice with OA or keyhole limpet haemocyanin in liposomes in a series of priming and boosting regimes failed to elicit any significant increase in serum or intestinal antibody response compared with feeding the free antigen. Oral tolerance induction to systemic challenge was also unaffected by OA entrapment in liposomes. In vitro liposome stability assays at 37 degrees C demonstrated a substantial resistance to disruption in the presence of acidic stomach contents. However, the addition of bile caused a rapid and profound release of protein marker from the liposomes. The rate and degree of disruption was influenced by the type of phospholipid used. These results suggest that liposomes may be useful as carriers for orally administered compounds but they are ineffective as adjuvants for the non-particulate, naturally weak immunogens used in this study.

Adjuvants, Immunologic↗

PubMed1992

The intestinal and serum humoral immune response of mice to orally administered antigens in liposomes: II. The response to liposome-entrapped bacterial proteins.

Effective oral adjuvants are needed to improve the intestinal immune responses to oral vaccines that are based on relatively low molecular weight antigens refined from veterinary pathogens. Liposomes prepared by different methods and composed of phospholipids of varying transition temperature were used to entrap cholera toxin (CT) and fed to mice. No significant increase in the intestinal antibody nor the serum IgA antibody response was detected but levels of serum IgG anti-CT antibody were significantly elevated in the group fed CT in phosphatidylcholine-based liposomes. Levels of antibody were significantly reduced in the groups fed CT in dipalmitoylphosphatidylcholine liposomes. Escherichia coli wall extract (ECWE) entrapped in certain liposome types and fed to mice elicited significantly increased serum anti-ECWE antibody responses but intestinal antibody responses were insignificantly different from the controls. These results suggest that orally administered liposomes fail to act as potent intestinal adjuvants for the entrapped antigens of bacterial origin used in this study.

Adjuvants, Immunologic↗

PubMed1992

Autoimmune haemolysis in the dog: relationship between anaemia and the levels of red blood cell bound immunoglobulins and complement measured by an enzyme-linked antiglobulin test.

A direct enzyme-linked antiglobulin test (DELAT) was used to measure the levels of red blood cell (RBC) bound IgG, IgM, IgA and C3 in dogs with autoimmune haemolytic anaemia (AIHA). At presentation, one or more DELAT parameters was raised in each AIHA case, and the RBC were typically coated with immunoglobulin of more than one class, together with C3. There was no relationship between the levels of RBC-bound IgG, IgM or IgA and the severity of the anaemia, although a significant negative correlation (rs = -0.66, P < 0.02) was found between bound C3 and blood haemoglobin concentration. These results indicate that the level of sensitisation of erythrocytes with IgG alone is not a reliable predictor of the severity of haemolysis in different cases, and that the pathogenesis of AIHA can be complex, involving multiple immunoglobulin classes and complement in the destruction of RBC. A significant relationship (rs = 0.63, P < 0.02) was found between serum IgG concentration and haemoglobin levels, and it is suggested that this may be due to free IgG inhibiting the interaction of IgG-sensitised RBC with macrophages. Serial measurements from individual AIHA cases during treatment revealed that the levels of RBC-bound immunoglobulins fell simultaneously with improvements in anaemia. In one dog, a relapse was associated with increases in bound IgG and IgM. Transient relative reticulocytopenia at presentation was common, but was not related to the severity of the anaemia. However, in other cases there was a persistent failure to increase RBC production, which was associated with slower recovery.

Anemia, Hemolytic, Autoimmune↗

PubMed1992

Depressed potential for interleukin-2 production following early weaning of piglets.

Spleen cells, but not mesenteric lymph node cells, from 3-week-old piglets abruptly weaned onto a soya-based diet, produced less interleukin-2 (IL-2) following non-specific activation with concanavalin A (Con A) than did cells from age- and litter-matched, unweaned controls. In contrast, the ability to express receptors for IL-2 was only marginally reduced. The effect on IL-2 production was most marked in animals weaned for as little as 24-48 h. Variation within groups increased with time after weaning, indicating differences between individuals in the longer-term effects of weaning. This finding may be due to endogenous production of steroids resulting in generalised impaired immune function or to retention of cells within intestinal sites owing to an active local immune response.

Animals↗

PubMed1992

Comparison of antibody and cell-mediated immune responses in horses following feeding of a novel dietary antigen, ovalbumin, and rotavirus.

Adult ponies which were fed ovalbumin (OVA) daily for 2 weeks had significantly greater serum anti-OVA IgG (P = 0.001) and antigen specific lymphocyte responses (P = 0.031) after intramuscular injection with OVA given with saponin than control ponies which had not been fed the antigen. This suggests that, despite the lack of evidence of B- or T-cell activation in peripheral blood during the period of OVA feeding, the animals were primed for an active secondary immune response. Adult ponies were challenged with equine rotavirus, strain H-2, but no statistically significant differences were found in serum IgG-associated antibody responses or antigen-specific lymphocyte responses between the rotavirus-challenged group and the control group, either following rotavirus challenge or intramuscular injection of rotavirus antigen given with saponin. Our findings, that feeding the non-replicating protein antigen OVA appeared to prime for an increased immune response rather than inducing oral tolerance, may be of relevance to future studies on the way the equine gastrointestinal tract handles usually harmless antigens.

Animals↗

PubMed1992

Impaired T-cell priming and proliferation in cats infected with feline immunodeficiency virus.

OBJECTIVE: Cats naturally infected with feline immunodeficiency virus (FIV) are particularly susceptible to infection with opportunistic pathogens, suggesting that these animals are unable to develop an effective immune response against the pathogen. Previous studies have used CD4+:CD8+ lymphocyte ratios and mitogen blastogenesis to identify immunological abnormalities in FIV-infected cats. However, these studies provide limited information for understanding the nature of the cellular dysfunction in FIV-infected cats, particularly defects in antigen-specific immune responses. DESIGN: To investigate whether cats infected with FIV are less able to mount an immune response to previously unencountered antigens, we compared the development of antigen-specific cellular immunity at the stage of T-cell priming in uninfected and FIV-infected cats. INTERVENTIONS: The general immune status of cats was assessed by peripheral blood CD4+:CD8+ lymphocyte ratios (flow cytometry), and by lymphocyte blastogenesis response to T- and B-cell mitogens. In addition, we describe the development of an autologous culture system to measure specific priming of naive feline T-cells to soluble antigen in vitro. This assay was used to compare T-cell priming in uninfected cats and cats which had been infected with FIV for 6-27 months. RESULTS: As in HIV infection, CD4+:CD8+ lymphocyte ratios in FIV-infected cats were found to be inverted, due to a reduction in the percentage of CD4+ cells. In addition, lymphocyte blastogenesis to both T- and B-cell mitogens was significantly impaired in FIV-infected cats. Priming to keyhole limpet haemocyanin (KLH) elicited a late proliferative response resulting from the expansion of CD4+ (T-helper cells). T-cell growth factor secretion correlated with cell proliferation. Restimulation of cells with fresh antigen-presenting cells and antigen showed that antigen-specific T-cell priming had occurred in the initial culture. When primary proliferation responses in FIV-infected cats were examined, it was observed that naive CD4+ T-cells from FIV-infected cats were significantly impaired (P less than 0.001) in their ability to be primed to KLH when compared with uninfected controls. CONCLUSIONS: Impaired priming of naive CD4+ T-helper cells to antigen in FIV-infected cats may explain the increased susceptibility of these animals to infection by opportunistic pathogens. The poor ability of human patients with AIDS to develop humoral immunity following vaccination may also be caused by such a defect in T-cell priming.

Acquired Immunodeficiency Syndrome↗

PubMed1992

An early defect in primary and secondary T cell responses in asymptomatic cats during acute feline immunodeficiency virus (FIV) infection.

As in HIV infection of humans, cats infected with FIV are particularly susceptible to secondary infection by opportunistic pathogens, suggesting an impaired ability to elicit an effective immune response against foreign antigens. In order to investigate the development of immunity in FIV-infected cats, we have used an autologous culture system to directly measure priming of naive CD4+ T cells to soluble protein antigen, in vitro. Using this assay, we showed previously that cats infected with FIV for several months had significantly reduced primary proliferative responses. We have now examined cats before infection, and at varying times after infection with FIV, to determine how soon after infection this defect in T cell priming was evident, compared with other quantitative and qualitative measurements of lymphocyte function. Our results showed a progressive decline in immune function in asymptomatic cats during the acute stage of infection with FIV. Primary T cell responses were most sensitive and a significant reduction in proliferation of naive T cells to foreign antigen occurred 5 weeks after infection, despite normal blastogenesis to T cell mitogens and normal CD4+/CD8+ ratios at these times. Whilst lymphocyte proliferation to T cell mitogens was unaffected throughout, a significant reduction in proliferation to a B cell mitogen occurred from week 8 onwards. CD4+/CD8+ ratios fell significantly from week 13 onwards, and proliferation of the memory T cell population to a recall antigen was significantly impaired later, from week 19 onwards. The defect in the priming of naive T cells to foreign antigen early after infection may be important in determining susceptibility to secondary infections.

Animals↗

PubMed1992

Immunocytological responses in porcine proliferative enteropathies.

The ileum, colon, and mesenteric lymph nodes of pigs naturally affected by either of the two major forms of proliferative enteropathy, namely, intestinal adenomatosis or hemorrhagic enteropathy, were examined for immunocytological responses to infection by immunocytochemistry, using antibodies directed against elements of the porcine immune system. In both forms, there was mucosal proliferation of immature enterocytes which lacked substantial major histocompatibility complex class II expression and a marked accumulation of immunoglobulin A (IgA) at the apical cytoplasm of affected enterocytes in association with intracellular Campylobacter-like organisms. In intestinal adenomatosis, there was only a mild infiltration of CD8+ and CD25+ T cells in the intestinal lamina propria. In hemorrhagic enteropathy, there was a moderate infiltration of CD8+ and CD25+ T cells and IgM+ B cells in the lamina propria. In rats and humans, villous enterocytes are thought to act as antigen-presenting cells, with major histocompatibility complex class II molecules present on their surface, capable of initiating a T-cell response (particularly of CD8+ T cells) in response to bacterial antigens. Therefore, the selection of immature crypt cells by the intracellular Campylobacter-like organisms for entry and multiplication may represent a remarkable microbial adaptation associated with local immunomodulation and enhanced bacterial survival. The accumulation of IgA within affected enterocytes may represent a reduced capability of the cells to process nonspecific IgA or an accumulation of specific IgA.

Animals↗

PubMed1992

Use of a direct enzyme-linked antiglobulin test for laboratory diagnosis of immune-mediated hemolytic anemia in dogs.

Detection of autoantibody, complement, or both bound to RBC is an essential requirement for unequivocal diagnosis of immune-mediated hemolytic anemia in dogs. An enzyme-linked antiglobulin test was adapted for laboratory diagnosis of this disease. The refinement and routine use of this assay have allowed further observation of the pathogenesis of the disease process. In particular, degree of hemolysis can be related to the degree of RBC sensitization associated with primary immune-mediated hemolytic anemia, and this correlation is highest for IgG autoantibody. Results indicate that autoantibody isotype might have an important role in the hemolytic process.

Anemia, Hemolytic, Autoimmune↗

PubMed1992

Primary antigen-specific T-cell proliferative responses following presentation of soluble protein antigen by cells from the murine small intestine.

To understand the local immune events which occur when a novel antigen is encountered in the gut it is necessary to know whether cells from the mucosal tissues are capable of initiating T-cell reactivity. We have examined the capacity of cells isolated from the Peyer's patches and the lamina propria of the murine small intestine to present keyhole limpet haemocyanin (KLH) to naive syngeneic splenic T cells in vitro. The properties of the gut antigen-presenting cells (APC) were compared with those of cells from the spleen and the mesenteric lymph nodes. Results clearly demonstrate that cells from the lamina propria as well as from the Peyer's patches, mesenteric lymph node and spleen can present KLH to naive T cells, inducing strong proliferative reactions comparable in magnitude and kinetics. All the APC populations tested induced interleukin-2 (IL-2) production in primary cultures, although minor differences were noted when lamina propria cells were used as APC. IL-4 was not detected in supernatants from cultures of non-immune T cells in the presence of APC from any tissue. Phenotypic analysis of the cells in cultures of naive T cells, with antigen and APC from different gut-associated tissues revealed important differences. Cells from Peyer's patch, mesenteric lymph node and spleen gave rise to cultures containing largely CD4+CD8- cells. However, cultures in which lamina propria cells acted as APC consisted primarily of CD4-CD8+ cells.

Animals↗

PubMed1991

The in vitro production of cytokines by mucosal lymphocytes immunized by oral administration of keyhole limpet hemocyanin using cholera toxin as an adjuvant.

The in vitro production of a variety of cytokines by lymphocytes isolated from spleen mesenteric lymph node (MLN), Peyer's patches (PP) and lamina propria (LP) was measured, after oral immunization with keyhole limpet hemocyanin using cholera toxin as an adjuvant. LP responses were characterized by very high levels of interleukin (IL) 4, IL 5 and IL 6 with lower levels of IL 2 and interferon-gamma (IFN-gamma). The PP had lower levels of IL 4, IL 5 and IL 6 than LP but higher levels of IL 2, IFN-gamma was only present at very low levels in this organ. The MLN had a pattern of cytokine production similar to the PP but did produce IFN-gamma. The spleen produced all cytokines measured except IL 5. Antibody production was characterized by IgA in the LP and PP but IgG was the dominant isotype in the spleen, the MLN was a poor source of antibody-producing cells. We interpret the results to show that (a) the LP response to cholera toxin/keyhole limpet hemocyanin is dominated by Th2-type cytokines compared to a lower production of Th1 type and (b) that the PP has responses typical of an organ with a high proportion of resting lymphocytes which develop mainly into Th2-types cells. The spleen is less dominated by Th2-type cytokines than the mucosal sites and this difference is paralleled by IgA antibody production at the mucosal sites and IgG antibody dominance in the spleen.

Adjuvants, Immunologic↗

PubMed1991

Immune responses to intramammary infusion with soluble (ovalbumin) and particulate (S uberis) antigens in the preparturient bovine udder.

Pregnant non-lactating cows were immunised by intramammary infusion with killed Streptococcus uberis into one quarter and ovalbumin into another, at one week (group 2) or one week and two weeks (group 1) before the expected date of parturition. A small IgG1 and IgG2 antibody response to ovalbumin was detected in the serum of these cows. There was also a small increase in IgG1 and IgA serum antibody activity to S uberis. In whey the response was restricted to IgA with activity to S uberis. The IgA antibody response to S uberis in group 1 was significantly greater in the quarter immunised with bacteria than that of the control quarters for up to two months after calving. In contrast, the serum IgA response was short or absent in a number of animals.

Animals↗

PubMed1991

Plasma and urine biochemical changes in cats with experimental immune complex glomerulonephritis.

Biochemical changes in plasma and urine were monitored in six cats before and during the induction of immune complex-mediated glomerulonephritis (ICGN) by daily intravenous administration of human serum albumin (HSA). The earliest indication of renal dysfunction in the cats was hypoalbuminaemia, which occurred as early as 13 weeks before cats developed clinical signs of renal disease. Proteinuria occurred 2 to 3 weeks before clinical disease, but was sensitive in predicting renal pathology in two cats that did not develop clinical signs of disease. In addition, increased activities of several urinary enzymes were detected in affected cats, with measurement of N-acetyl-beta-D-glucosaminidase and gamma-glutamyl transferase providing the earliest and most sensitive indication of renal damage. These plasma and urine measurements correlated more closely with the renal pathology, observed at postmortem, than clinical assessment of disease. It was concluded that ICGN in the cat could be diagnosed earliest by measurement of plasma protein concentration, whilst disease progress could be effectively monitored by including assays to measure urine protein and urine enzymes.

Animals↗

PubMed1991

Identification of autoantigens in canine autoimmune haemolytic anaemia.

Autoantigens in canine autoimmune haemolytic anaemia (AIHA) were identified by immunoprecipitation using autoantibody eluted from the erythrocytes of affected dogs. At least three patterns of precipitated antigen were identified in six cases of AIHA. The most commonly precipitated antigen pattern was a combination of 42-kD and 29-kD peptides, associated with up to three other membrane components. These autoantigens may be canine glycophorins, which are of similar molecular mass, or may possibly represent an equivalent of the human Rhesus complex. An autoantigen identical in molecular mass to band 3, the erythrocyte anion channel protein, was precipitated in one case of AIHA, and unknown peptides of 37 kD and 100 kD were isolated by autoantibody from another dog. In one case, no antigens were precipitated by the eluted antibody, indicating that the autoantibody may have bound a non-protein membrane component such as phospholipid. Overall it is considered that the different patterns observed may reflect differences in the aetiology of the condition. In other studies, sera from dogs with AIHA failed to immunoprecipitate autoantigens, but were shown by immunoblotting to contain autoantibodies to proteins of the erythrocyte cytoskeleton. Such autoantibodies were also demonstrated in normal canine sera and it is suggested that they are unlikely to play a role in the pathogenesis of AIHA, but may be part of a normal clearance mechanism for damaged red blood cells.

Anemia, Hemolytic, Autoimmune↗

PubMed1991

Dietary antigen handling by mother and offspring.

Sows were fed ovalbumin (OvA) as a novel protein antigen either throughout gestation and lactation (G + L) or during lactation only (L). This resulted in a significant uptake of OvA into blood, colostrum and milk along with a specific IgG response. In piglets from the G + L group, OvA and antibodies to OvA were detected in serum after ingestion of colostrum. In a large proportion of these piglets OvA was still detected at 3 weeks of age. In the L group a significant proportion of the piglets responded to OvA whilst still suckling their mother. At 3 weeks of age all piglets were weaned onto an egg-based diet. A similar uptake of OvA was seen in all piglets but there was no response to OvA in the G + L piglets. In piglets from sows fed only during lactation, however, a rapid IgG anti-OvA response and signs of diarrhoea were seen. The results suggest that factors of immunological importance are passed over from mother to offspring and it is proposed that immunological experience of dietary antigens by the mother is important for a 'safe' tolerance induction in her offspring.

Animals↗