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Biomedical subjects

J D Reynolds

Publications and source records attributed to J D Reynolds.

At least 19 recordsLinked to original sources
PubMed1995

Apoptosis of ileal Peyer's patch B cells is increased by glucocorticoids or anti-immunoglobulin antibodies.

The ileal Peyer's patch (PP) in sheep plays a central role in the development and production of B cells. Associated with a tremendous amount of B cell proliferation in this site is the extensive diversification of the Ig repertoire by somatic hypermutation. Very few (< 5%) of the B cells produced in the ileal PP differentiate and emigrate; instead, the vast majority of these cells soon die, and we have previously shown that death is associated with apoptosis. When placed in culture, ileal PP B cells die rapidly by apoptosis, such that after 24 h, 60 +/- 1% of DNA is fragmented. Here, we show that the extent of this spontaneous B cell apoptosis in culture, as quantitated by DNA fragmentation, was significantly increased in a dose-dependent manner by the glucocorticoids hydrocortisone or dexamethasone. Furthermore, treatment of lambs with 2-2.5 mg/kg of dexamethasone resulted in a marked increase in the number of apoptotic cells in the ileal PP and an increase in ileal PP B cell DNA fragmentation to 20 +/- 6%, compared with 2.4 +/- 0.1% in untreated lambs. Anti-immunoglobulin (Ig) antibodies also increased the extent of DNA fragmentation in cultured ileal PP B cells. After 24 or 48 h of culture with anti-Ig (PIg47A), DNA fragmentation was 74 +/- 2% and 75 +/- 3%, respectively. Ileal PP B cells are rescued from apoptosis by agents that activate protein kinase C and increase cytosolic Ca2+, and here we show that this treatment also results in apoptotic rescue in the presence of dexamethasone or anti-Ig. We speculate that the apoptosis of ileal PP B cells in situ may be modulated by glucocorticoids and by the cross-linking of surface Ig. Apoptosis, induced by a signal through surface Ig, may be an important mechanism in the deletion of self-reactive B cells during the expansion of the Ig repertoire in the ileal PP.

Animals↗
PubMed1995

Rescue of ileal Peyer's patch B cells from apoptosis is associated with the induction of Bcl-2 expression.

The major site of B-cell genesis in the sheep is the ileal Peyer's patch (PP). The B cells in the ileal PP undergo both extensive proliferation and massive death in association with an ongoing diversification of the immunoglobulin repertoire by somatic hypermutation. Most, if not all, the B-cell death in the ileal PP is due to apoptosis. When placed in culture, ileal PP B cells undergo rapid apoptosis. Here, we investigated the expression of the proto-oncogene bcl-2 in ileal PP cells in situ and in culture. Bcl-2 expression has been correlated with the prevention of apoptosis in many cell types. Western blotting, using anti-Bcl-2 monoclonal antibodies, revealed that a Bcl-2-reactive protein of 26,000 MW was expressed in ileal mesenteric lymph node cells, splenocytes and thymocytes from sheep, but was barely detectable in ileal PP B cells in situ or in culture. However, Bcl-2 expression could be markedly induced in ileal PP B cells cultured with phorbol ester and Ca2+ ionophore, a procedure that is known to rescue these cells from apoptosis. We hypothesize that those few B cells that survive a selection event in the ileal PP may begin to express elevated levels of Bcl-2 as they escape from the apoptotic pathway.

Animals↗
PubMed1995

Comparison of recognition and grating acuities in very-low-birth-weight children with and without retinal residua of retinopathy of prematurity. Cryotherapy for Retinopathy of Prematurity Cooperative Group.

PURPOSE: To compare recognition (letter) and resolution (grating) acuity in eyes with and without retinopathy of prematurity (ROP) residua. METHOD: Letter acuity measured with the crowded HOTV chart or pocket cards (Good-Lite), and grating acuity measured with the Teller Acuity Card (TAC) procedure, were compared at the 3 1/2- and 4 1/2-year follow-up exams in the CRYO-ROP study. Testers were unaware of the retinal status of individual eyes. RESULTS: Measurable scores for both letter and grating acuity were obtained from 1694 eyes at 3 1/2 years and 2101 eyes at 4 1/2 years. Correlation analysis showed that the relation between crowded HOTV and grating acuity scores was best described by a quadratic function, with r2 values of 0.57 at 3 1/2 years and 0.68 at 4 1/2 years. Difference scores were calculated for each eye by subtracting the log of the HOTV score (converted to cyc/deg based on the convention that 20/20 = 30 cyc/deg) from the log TAC score. Normal eyes showed HOTV acuity that was higher than TAC acuity by an average of 0.37 octave (SD = 0.46, n = 1150) at 3 1/2 years and 0.27 octave (SD = 0.43, n = 1337) at 4 1/2 years. HOTV and grating acuity scores were similar to each other in eyes with retinal residua of ROP. Overall, eyes with acuity better than 20/150 to 20/300 tended to show better HOTV than grating acuity, whereas those with acuity below 20/150 to 20/300 generally showed better grating than HOTV acuity. CONCLUSIONS: There is a significant correlation between crowded HOTV letter acuity and TAC grating acuity in young children for normal eyes and eyes with ROP residua. In both groups of eyes, eyes with lower acuity show better grating than letter acuity, whereas eyes with better acuity show slightly higher letter than grating acuity.

Child, Preschool↗
PubMed1992

Ontogeny of glutamate and gamma-aminobutyric acid release in the hippocampus of the guinea pig.

The ontogeny of the L-glutamate (GLU) and gamma-aminobutyric acid (GABA) neuronal systems in the guinea pig hippocampus was investigated with respect to tissue amino acid content, and spontaneous and K(+)-stimulated release of GLU and GABA. Transverse hippocampal slices were prepared from the guinea pig fetus at day 45 (brain growth spurt), 55 and 63 of gestation (term, about 68 days), from the 5-days-old neonate, and from the young adult. GLU and GABA release was determined as efflux from hippocampal slices into Krebs'-bicarbonate medium using a dynamic, submerged, superfusion apparatus. Hippocampal GLU content decreased during development, whereas GABA content was constant for all the ages investigated. The magnitude of spontaneous GLU efflux decreased during development; there was no measurable spontaneous GABA efflux. The K+ concentration-GLU efflux response curve was bell-shaped for the fetus at the three selected gestational ages, and was curvilinear for the neonate and adult. The apparent EC75 of K(+)-stimulated GLU efflux was higher for the neonate and adult compared with the fetus. In contrast, the K+ concentration-GABA efflux response curve was curvilinear, and the apparent EC75 of K+ was similar for all the ages investigated. K(+)-stimulated efflux of GLU and GABA was Ca++ dependent, but this was not the case for spontaneous GLU efflux. These data indicate that, in the guinea pig hippocampus, the GLU neuronal system is developing throughout gestation, whereas the GABA neuronal system appears to mature before the brain growth spurt.

Animals↗
PubMed1992

T-cell receptor-gamma delta association with lymphocyte populations in sheep intestinal mucosa.

T cells expressing T-cell receptor (TcR)-gamma delta and CD8 represent a significant population in mouse and chicken intra-epithelial lymphocytes (IEL) but represent a minor population in human IEL. We examined the TcR-gamma delta usage and co-expression of CD5, CD4, CD8 and major histocompatibility complex (MHC) class II on isolated sheep IEL and lamina propria lymphocytes (LPL), and compared them with the TcR-gamma delta + cells in peripheral blood, intestinal lymph and jejunal Peyer's patches (PP). There were a number of notable differences. TcR-gamma delta + cells comprised 18% of IEL and 10% of LPL. Among the population of TcR-gamma delta + IEL, 24% were CD8+ and 54% were CD5+, which contrasts with the TcR-gamma delta + cells in blood and intestinal lymph that were universally CD5+ CD4- CD8-. A notable feature of the IEL was the presence of distinct CD8+ and TcR-gamma delta + populations that lacked CD5. Also a high percentage of IEL and LPL were CD2+ and MHC class II+. Analysis of the expression of MHC class II on T-cell subsets, as an indicator of activation, showed that 60-95% of the various IEL and LPL subsets were MHC class II+ compared with only 5-40% in jejunal PP, lymph nodes, spleen and blood. Therefore, it is possible that the circulating TcR-gamma delta + and CD8+ cells that localize in the gut epithelium might become activated and stop the expression of CD5 under the influence of the local microenvironment. These cells appear not to emigrate while still expressing the TcR-gamma delta + (CD8+) CD5- MHC class II+ phenotype. Our data, together with those from other studies, show that there is much heterogeneity in the use of TcR-gamma delta and accessory T-cell molecules by IEL.

Animals↗
PubMed1992

Characterization of B-cell phenotypic changes during ileal and jejunal Peyer's patch development in sheep.

Changes in B-cell phenotype during development of ileal and jejunal Peyer's patches (PP) of sheep were investigated using flow cytometry and immunoperoxidase-stained cryosections. On Day 104 of gestation (term at 150 days) B-cell clusters were identified in the lamina propria of the ileum. These clusters were composed of cells that expressed surface IgM (sIgM), lambda or kappa light chain, and BAQ44A, a B-cell differentiation molecule. No cells in the clusters stained for terminal deoxynucleotide transferase. On Day 132 gestation, a change was evident in the phenotype of ileal PP B cells. Most B cells expressed a reduced level of sIgM and 20% were BAQ44A-. The B cells in the dome region were BAQ44A+ but few BAQ44A+ cells were present in the follicles. At 6-8 weeks of age BAQ44A+ cells were restricted to the dome region of the ileal PP; flow cytometric analysis confirmed that 25% of B cells isolated from the dome/follicle complex were BAQ44A+. Thus, the primordial PP was populated with B cells that were phenotypically similar to circulating B cells (sIgMhigh, BAQ44A+). After 132 days gestation, the predominant B-cell phenotype in the ileal PP changed to sIgMlow and BAQ44A-. This phenotypic change could be the result of either early immigrant B-cell differentiation or subsequent colonization by sIgMlow BAQ44A- B cells. The phenotypic changes of ileal PP follicular B cells were not complete until after birth and different phenotypic changes were observed in follicles of the jejunal PP of young lambs.

Animals↗
PubMed1992

An analysis of the growth and differentiation of B cells isolated from follicles of the ileal Peyer's patch of sheep.

We developed a method to isolate and culture cells from the lymphoid follicles of the ileal Peyer's (PP) patch of young sheep (6-12 weeks). These cells were 98% sIgM+ B cells and 1% T cells. Cultured follicular cells were used to investigate B-cell proliferation and differentiation. Less than 50% of B cells were viable after 24 hr of culture and this decrease in B-cell viability also occurred following co-stimulation with pokeweed mitogen (PWM) and recombinant bovine interleukin-1 (rBoIL-1) or rBoIL-2. In contrast, co-stimulation with PWM and either rBoIL-1 or rBoIL-2 induced a marked proliferative response that was maximal on Day 4 of culture. Cytokine-induced proliferation of the B cells required PWM co-stimulation and proliferation induced by rBoIL-1 and rBoIL-2 was neither additive or synergistic. This suggests that PWM bound a molecule or molecules that signalled responsiveness to both rBoIL-1 and rBoIL-2. Culture of follicular cells with PWM and both rBoIL-1 and rBoIL-2 also resulted in B-cell differentiation. This differentiation was associated with decreased proliferation, an increased number of viable B cells, and increased expression of both surface IgM and non-Ig membrane molecules. Thus, co-stimulation of ileal PP follicular cells with PWM and rBoIL-1 and rBoIL-2 resulted in both B-cell proliferation and differentiation.

Animals↗
PubMed1991

Capsaicin effects on non-neuronal plasma membranes.

Capsaicin has been touted as a pharmacological tool specific for sensory afferent neurons and is widely used in neurophysiological studies. However, we have recently demonstrated that in concentrations commonly employed within the gastrointestinal tract, capsaicin inhibits platelet aggregation to at least three different stimuli. Since this was observed in a nerve free system it raised the question of how specific capsaicin is. In this communication we report that capsaicin has profound effects on physical properties of non-neuronal cell plasma membranes. These effects were observed while measuring the effect of capsaicin upon the fluidity of both intact cell membranes and a variety of purified membrane preparations. Membrane fluidity was assessed with the fluorescent probes diphenylhexatriene (DPH) and its trimethylamino derivative TMA-DPH and demonstrated concentration-dependent capsaicin effects. Furthermore, the effects were cell specific and for full expression required both intact cells and a non-lipid extractable component of the plasma membrane. These non-neuronal effects must be carefully considered when contemplating the explanation for capsaicin-induced effects.

Animals↗
PubMed1991

Ileal Peyer's patch emigrants are predominantly B cells and travel to all lymphoid tissues in sheep.

The ileal Peyer's patch (PP) was selectively labeled with fluorescein isothiocyanate by extracorporeal perfusion in 7-12 week-old lambs and the lymphocyte lineage and fate of the emigrants was determined by fluorescence microscopy and flow cytometry. PP emigrants were found in all tissues examined, accounting for 10%-15% of ileal mesenteric lymph node (MLN). 1%-2% of jejunal MLN, jejunal PP, prescapular lymph node (PLN) and 3%-4% of spleen cells. All ileal PP emigrants enter the ileal MLN on their way to the circulation. Removal of the MLN prior to perfusion enabled emigrants to go directly to the circulation and extravasate in distant tissues faster than in intact animals. The ileal MLN might provide an additional level of regulation for ileal PP emigrants. The perfused ileal PP contained about 25 times more B cells than T cells. The emigrant cells found in different tissues included both T and B cells but came to reflect, although to a lesser degree, the B cell composition of the tissue from which they were derived. One day after perfusion the composition of PP emigrants was similar to that of the tissue within which they were found; the spleen was the exception with a bias towards B cells. By day 3 the ratio of B to T cells in the PP emigrants was 1 for jejunal MLN and PLN. 1.5 for ileal MLN and jejunal PP, and 4-5 for the spleen and blood. It was concluded that the PP-derived T cells were recirculating T cells that were in the ileal PP at the time of perfusion. These cells emigrated rapidly and equilibrated such that they accounted for about 1.5% of the T cell pool in various tissues. Most PP-derived B cells were probably produced in the PP. The greatest contribution (24.4%) that ileal PP emigrants made to the B cell pool of a tissue was with the ileal MLN through which they are obliged to pass. The contribution was lower but still very significant in blood (8.9%), spleen (6.8%), PLN (3.9%), jejunal MLN (3.5%) and jejunal PP (1.8%). There was no evidence that ileal PP emigrants made a greater relative contribution to either T or B cell populations in MLN or jejunal PP than to non-gut-associated sites. The B cells were distributed throughout the immune system, which is in accordance with the proposal that the ileal PP is a site of primary B cell genesis in sheep.

Animals↗
PubMed1991

Apoptosis is associated with the extensive B cell death in the sheep ileal Peyer's patch and the chicken bursa of Fabricius: a possible role in B cell selection.

The ileal Peyer's patch (PP) and the bursa of Fabricius have major roles in populating the B cell system in sheep and chickens, respectively. These tissues contain greater than 90% B cells and possess a massive proliferation index with greater than 5% of B cells entering mitosis per hour. Paradoxically, almost all of the B cells produced in these sites rapidly die in situ. Here we show that the extensive B cell death occurring in the ileal PP and bursa is associated with apoptosis. Gel electrophoresis of ileal PP cell DNA from 7-14 week-old lambs and bursal cell DNA from 4-week-old chickens demonstrated a laddering of DNA in multiples of approximately 200 bp, a pattern indicative of apoptosis. In sheep, the intensity of the laddering pattern seen after agarose gel electrophoresis was always greater with ileal PP cell DNA compared with thymocyte DNA, and usually greater than jejunal PP cell DNA. Likewise, DNA isolated from chicken bursal cells and mouse PP cells always exhibited a more intense laddering pattern than chicken or mouse thymocytes, respectively. When placed in culture ileal PP cells died rapidly less than 40% viable cells were recovered after 24 h. Within 6 h of culture many ileal PP cells exhibited an apoptotic appearance in that they contained condensed chromatin and fragmented nuclei. Moreover, greater than 55% of total cellular DNA was fragmented. Compared with thymocytes, ileal PP cells underwent DNA fragmentation to a much greater extent and with a faster time course in short-term culture. We propose that cell death by apoptosis may make an important contribution to B cell development in the lamb ileal PP and the chicken bursa. Apoptosis may provide a mechanism for the diversification of the B cell immune repertoire and/or the selection of non-self reactive B cells.

Animals↗
PubMed1991

Negative signaling by surface IgM on B cells isolated from ileal Peyer's patch follicles of sheep.

Lymphoid follicles from the sheep ileal Peyer's patch (PP) were used to prepare a cell suspension consisting of 98% surface IgM-positive (sIgM+) B cells and 1% T cells. Co-stimulation of follicular cells with pokeweed mitogen and either recombinant bovine interleukin 1 (IL 1) or IL 2 resulted in a marked proliferative response. In contrast, the addition of soluble F(ab')2 rabbit anti-sheep Ig completely inhibited the proliferative response induced by pokeweed mitogen and IL 1 or IL 2 co-stimulation. Anti-Ig inhibition of B cell proliferation was specific for ileal PP follicular cells and was not observed with mesenteric lymph node cells or splenocytes. Furthermore, suppression of ileal PP follicular B cell proliferation required at most divalent cross-linking of sIg was independent of Fc receptors, but was dependent on the concentration of anti-Ig and required 48 h for maximal effect. Negative signaling by sIgM indicates that ileal PP follicular B cells are functionally distinct from B cells in other secondary lymphoid tissues. Also, the present observations are consistent with previous reports indicating that B cell proliferation in ileal PP follicles is antigen independent.

Animals↗
PubMed1991

Evaluation of edetate and thiamine for treatment of experimentally induced environmental lead poisoning in cattle.

Twenty mature Holstein cows were randomized into 5 treatment groups. Cows of groups 2 to 5 were given 2 mg of elemental Pb/kg of body weight for 28 days. Clinical signs of plumbism were scored, and blood for Pb, progesterone, and hematologic analyses was collected weekly. Cows also were examined weekly for anomalous ovarian cycles. Starting on study day 28, cows in group 3 were treated once daily with 2 mg of thiamine HCl/kg (IM) for 13 days, cows in group 4 were treated twice daily with 62 mg of Na2,Ca-EDTA/kg (IV) for 4 days, and cows in group 5 were given thiamine (dosage regimen the same as for group 3) plus Na2,Ca-EDTA (dosage regimen the same as for group 4). On study days 96 through 139, cows were slaughtered in a commercial abattoir and samples of blood, skeletal muscles, bones, liver, and kidneys were collected and assayed for Pb concentration. Thiamine was not effective in reducing blood Pb concentration, and treatment with Na2,Ca-EDTA and thiamine plus Na2,Ca-EDTA was effective in reducing the concentration of Pb in blood. However, treatment with thiamine was more effective than treatment with Na2,Ca-EDTA or thiamine plus Na2,Ca-EDTA in inducing remission of clinical signs of plumbism. The concentration of Pb in blood was significantly (P less than 0.05) correlated to the concentration of Pb in liver, kidneys, skeletal muscles, and bones. Significant (P less than 0.05) relationship existed between number of days from Pb exposure to slaughter and concentration of Pb in blood, liver, and skeletal muscles. Exposure to Pb did not significantly alter CBC values.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1990

Observations on PCBs and mercury in common loons (Gavia immer) collected from southwestern Lake Michigan.

The death of common loons (Gavia immer) was associated with a small spill of bunker-C oil off the Chicago shoreline of Lake Michigan. Petroleum oil was not found on the feathers or in the lungs of the birds. Botulinus toxins C and E were found in heart blood. Because the carcasses were autolysed, botulism toxins could have been produced postmortem. An average of 97 micrograms PCBs (Aroclor 1254 standard) and 2.2 micrograms dieldrin/g of body fat also were found. Concentrations of heavy metals in one bird were 0.25 microgram of total mercury and 0.5 microgram of lead/g of liver, respectively. The loons had abundant body fat suggesting they were not debilitated at the time of death.

Animals↗
PubMed1989

Two types of sheep Peyer's patches: location along gut does not influence involution.

In sheep, and some other species, there is evidence of two types of Peyer's patches (PPs), the ileal PP, which extends 150 cm along the terminal ileum, and the jejunal PPs distributed throughout the rest of the small intestine. The two types differ significantly in their histology, ontogeny and the extent of lymphocyte traffic. Another intriguing difference is that the ileal PP involutes about the time of puberty whereas the jejunal PPs function throughout life. This study shows that the differences in PP lifespan is not related to their specific location in the small intestine. Surgery was done at 1-2 months of age to transpose lengths of ileal PP into the jejunum, also, PP-containing lengths of jejunum were transposed into the midst of the ileal PP. Examination at age 12-16 months showed that ileal PP transposed into jejunum had involuted at the same rate as normally sited ileal PP. Also, jejunal PPs transposed into the ileum had not involuted unlike the surrounding ileal PP. It was concluded that the difference in lifespan of the two PPs were not related to the local microenvironment created by gut function, but may be inherent to the PP itself.

Aging↗
PubMed1988

T and B cells have similar recirculation kinetics in sheep.

T and B cells continually migrate around the body by recirculating between blood and lymph via lymphoid tissues. This is an important way of fostering the cell-cell interactions required for the initiation of an immune response. Data from previous studies of T and B cell recirculation, based on relatively complex approaches, have been interpreted as evidence that the recirculation of B cells is more sluggish than for T cells. We have shown from a study in sheep that this is not the case. Lymphocyte migration through both the intestine and the prescapular lymph node was analyzed by cannulating efferent lymphatics. The lymph cells were labeled in vitro with Hoechst 33342, or other fluorochromes, then injected i.v., and the rate of their recirculation into the lymph was determined by fluorescence microscopy. A multicolor immunofluorescence assay was used to classify each recirculated cell as either a T or B cell. This established that the T and B cells from intestinal lymph and from prescapular lymph recirculate with similar kinetics. The recovery of i.v. injected T and B cells was the same in prescapular lymph, but fewer injected B cells than T cells were recovered in intestinal lymph. Thus, although the recirculation kinetics are the same for T and B cells, the two populations are handled differently in intestinal tissues and a lymph node remote from the gut.

Animals↗
PubMed1987

Peyer's patches export lymphocytes throughout the lymphoid system in sheep.

The lymphocyte output from small intestine containing either the long continuous ileal Peyer's patch (PP) or several smaller jejunal PP was examined in young lambs. Most studies were done in 2-mo-old lambs, 1 mo after removal of mesenteric lymph nodes (MLN). Extracorporeal perfusion of part of the intestine and addition of fluorescein isothiocyanate to the perfusate led to the labeling, in their normal microenvironment, of a regionally defined population of cells. One day later considerable numbers of emigrant lymphocytes were identified by fluorescence microscopy in the spleen, MLN and peripheral lymph nodes, jejunal PP, and bone marrow. In nonperfused ileal PP and thymus the labeling indexes were low. The highest labeling index was in the blood where 3.7% of the lymphocytes were labeled. A similar organ distribution of emigrant cells was found on day 3. When MLN were included in the perfused region more emigrants were identified. In some animals the intestinal lymphatic draining the perfused ileum was cannulated. Continual lymph drainage caused a dramatic decrease in the labeling indexes in other lymphoid organs. A substantial number of lymphocytes leave both ileal PP and jejunal PP via lymphatics and travel to all other lymphoid organs. However, the number of emigrant lymphocytes compared with the total number of labeled lymphocytes in the perfused tissue was about 10 times greater after perfusing gut with the jejunal PP than after ileal PP perfusion. We conclude that relatively more lymphocytes emigrate from the jejunal PP than from the ileal PP.

Animals↗