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Biomedical subjects

B J Gormus

Publications and source records attributed to B J Gormus.

At least 55 records · Page 3Linked to original sources

Experimental leprosy in the mangabey (Cercocebus atys): necropsy findings.

A mangabey monkey (Cercocebus atys) was inoculated intravenously and intracutaneously with acid-fast bacilli (AFB) from a mangabey with spontaneously acquired leprosy. It developed generalized lepromatous leprosy and died 46 months after inoculation. Necropsy revealed severe lepromatous infiltrates in the skin, nasal mucosa, peripheral nerves, and testicles. Internal organs were only minimally involved. The lesions seen at necropsy were very similar to those seen in untreated cases of human lepromatous leprosy. These findings further substantiate the mangabey monkey as a suitable animal model for the study of lepromatous leprosy.

Animals↗

Electron microscopic study of leprosy in a mangabey monkey (natural infection).

Ultrastructural features of the leproma of a) a naturally infected mangabey monkey, and lepromas and liver of b) a passage mangabey monkey, c) a rhesus monkey, d) an African green monkey, and e) a nine-banded armadillo inoculated with leprosy bacilli isolated from the leproma of a naturally infected mangabey monkey were studied by the freeze-etching technique. The size, shape, and ultrastructural features of leprosy bacilli in the phagolysosomes of macrophages in all of these samples were essentially the same as those in humans, nude mice, and armadillos inoculated with human Mycobacterium leprae. Distinct accumulations of small spherical droplets were observed around leprosy bacilli inside lepra cells of all the samples but were scarce in the specimen from the green monkey. The bacilli in all samples were long and slender, and had band structures on the smooth cell wall surfaces. The bacilli were indistinguishable from M. leprae.

Animals↗

Functional analysis of monkey lymphocyte subsets defined by OKT4 and OKT8 Monoclonal Antibodies.

The percentages of rhesus monkey blood lymphocytes (PBL) reactive with OKT4 and OKT8 antibodies and the OKT4/OKT8 ratio showed significant correlations with the log of the immunoglobulin plaque-forming cell (PFC) response after stimulation with pokeweed mitogen (PWM). These correlations suggested that monkey OKT4+ cells function as "helper" cells and OKT8+ cells function as "suppressor" cells for the PFC response. This was confirmed by separation and study of enriched T- and B-cell subpopulations. OKT8-depleted (OKT4+) and OKT4-depleted (OKT8+) cells were obtained by treatment of purified T cells with antibody and complement. OKT4+ cells augmented the PWM-induced B-cell differentiation into PFC but OKT8+ cells did not. OKT8+ cells suppressed the PFC response by mixtures of B cells and OKT4+ cells. OKT8 antibodies also detected a suppressive cell subset in African green monkeys since the percentage of OKT8+ cells showed a negative correlation with the log PFC response. OKT4 antibodies failed to bind to African green monkey PBL.

Animals↗

Heterogeneity of human lymphocyte Fc receptors: studies using heat-aggregated and antigen-complexed IgG from human, rabbit, guinea pig, horse and goat.

We have studied the ability of human peripheral blood lymphocytes (HuPBL)4 to interact with IgG from several animal species. Three functions or activities that are reported to depend on an interaction between complexed IgG and HuPBL receptors (R) for the Fc piece of IgG (Fc gamma R) were compared: (1) antibody-dependent cell-mediated cytotoxicity (ADCC); (2) binding of heat-aggregated IgG (aggG); and (3) rosette formation with IgG-sensitized erythrocytes [RBC-A(gamma)]. IgG (and IgM) antibodies to chicken erythrocytes (CRBC) were purified from the sera of the following species after injection with CRBC stroma: (1) horse (Ho); (2) goat (Go); (3) rabbit (Ra); and (4) guinea pig (Gp). Good IgG-agglutinating antibody titers were obtained from each injected species. Using 51Cr-labeled CRBC targets and HuPBL effector cells, only Ra anti-CRBC IgG gave good ADCC at high dilutions. Ho and Go anti-CRBC (IgG) failed to give ADCC, and Gp anti-CRBC (IgG) gave approx. 30% of the level of kill as Ra. Ra Fab2 fragments of IgG antibody failed to produce ADCC. Treatment of HuPBL with Ra anti-lymphocyte serum (ALS) almost totally ablated ADCC, whereas HoALS failed to alter ADCC. Pretreatment of HuPBL with aggG showed that Ra or Hu aggG gave essentially equal inhibition of ADCC, Gp gave approx. 30% of the degree of inhibition as Hu and Ra, and Ho or Go aggG had essentially no effect of ADCC. These results confirmed the following order of ability of IgG to interact with HuPBL ADCC killer (K) cells: (Hu greater than or equal to)Ra greater than Gp much greater than Ho, Go. The data suggest that Gp IgG interacts with only a subpopulation (approximately 30%) of HuPBL K cells. The binding of aggG to total HuPBL failed to strictly correlate with the ADCC results or with the results of rosette formation between total HuPBL and CRBC-A(gamma). The observations suggest that there is a heterogeneity of Fc gamma R between K and non-K cell subpopulations of HuPBL both in terms of the type of complexed IgG they are able to bind, and in terms of species of origin of the IgG. The data also support contentions that Fc gamma R that bind RBC-A (gamma) complexes differ from those that bind aggG.

Animals↗

Capping of peripheral blood lymphocyte C3 receptors in chronic lymphocytic leukemia: relationship to chemotherapy.

The data show that in chronic lymphocytic leukemia (CLL) there are elevated numbers of peripheral blood lymphocytes (PBL) capable of rosette formation (complement-rosetting lymphocytes, CRL) with bacteria (B)-antibody-(A)-complement (C) (BAC) complexes. In most of the CLL patients studied, large percentages of CRL were impaired in their ability to undergo BAC-C3R cap formation compared to normal donors. The degree of impairment varied from one patient to the next. A consideration of clinical information suggested that the variation was related to the clinical response of patients to chemotherapy. Four categories of CLL patients could be identified on the basis of clinical data: (1) patients never received therapy for CLL; (2) those whose clinical history indicated a failure to respond to ongoing chemotherapy; (3) patients who had received successful prior chemotherapy and who were asymptomatic or stable and were receiving no therapy at the time of study; and (4) those who were undergoing therapy at the time of study and who subsequently became stable or asymptomatic and were removed from therapy. The first two categories of patients failed to show significant BAC-C3R capping, whereas the last two were able to form C3R caps on significant percentages of CRL. No direct relationship could be discerned between C3R capping and currently accepted classifications on stages (Rai) of CLL. These results suggest that monitoring the percentages of BAC-C3R cap-forming CRL may be useful in determining the success of chemotherapy of CLL patients. The microtubule-disrupting agent colchicine enhanced BAC-C3R capping in all CLL patients to normal levels, whereas the microfilament-disrupting agents cytochalasin B(CB) had no effect on CLL RBL C3R capping. CB totally abolished C3R capping of CRL from normal donors. These results suggest that abnormalities exist in both cytoskeletal and contractile systems of CR from CLL peripheral blood.

Antineoplastic Agents↗

Human peripheral blood erythroid burst forming unit (BFU(E)): evidence against T-lymphocyte requirement for proliferation in vitro.

Erythroid burst-forming units (BFU(E)) are proliferative cells which may be precursors of the erythroid colony-forming unit (CFU(E)). To examine the role of T lymphocytes in the proliferation and/or differentiation of human blood BFU(E), the effect of purified T lymphocytes on erythroid colony (EC) formation by purified null cells was examined in vitro. Lymphocyte subpopulations were prepared by Ficoll-Hypaque centrifugation, immunoadsorbent chromatography, and sheep red blood cell rosetting after removal of monocytes by adherence to plastic. Cultures of isolated B, T, or null lymphocytes alone revealed that BFU(E) were present in the null cell fraction. Addition of isolated B and/or T lymphocytes in various ratios to null cells failed to influence the number or size of EC formed. These results indicate that normal human circulating BFU(E) are contained in the null cell fraction of peripheral blood lymphocytes and do not require T lymphocytes for normal growth and differentiation in vitro.

B-Lymphocytes↗

Heterogeneity among human lymphocyte effector cells mediating spontaneous lymphocyte-mediated cytotoxicity.

The spontaneous lymphocyte-mediated cytotoxicity (SLMC) of lymphocytes from normal individuals was studied using the lymphocyte titration assay. Various techniques were used to study selectively several lymphocyte subpopulations. The activity of these subpopulations was tested in both the antibody-dependent cellular cytotoxicity (ADCC) and lymphocyte titration assays. Surface marker studies were performed on each lymphocyte subpopulation. The results from these studies indicate that effector cells mediating SLMC are a heterogeneous lymphocyte population composed of Fc-receptor-positive lymphocytes both with and without C3 receptors that consistently segregate with cells that do not form rosettes with sheep red blood cells. Our data suggest that, in many respects, the effector cells in SLMC and ADCC are identical. However, the mechanism appears to be different.

Adult↗

Cell-cell interaction in erythropoiesis. Role of human monocytes.

Erythroid burst forming units (BFU-E) are proliferative cells present in peripheral blood and bone marrow which may be precursors of the erythroid colony forming cell found in the bone marrow. To examine the possible role of monocyte-macrophages in the modulation of erythropoiesis, the effect of monocytes on peripheral blood BFU-E proliferation in response to erythropoietin was investigated in the plasma clot culture system. Peripheral blood mononuclear cells from normal human donors were separated into four fractions. Fraction-I cells were obtained from the interface of Ficoll-Hypaque gradients (20-30% monocytes; 60-80% lymphocytes); fraction-II cells were fraction-I cells that were nonadherent to plastic (2-10% monocytes; 90-98% lymphocytes); fraction-III cells were obtained by incubation of fraction-II cells with carbonyl iron followed by Ficoll-Hypaque centrifugation (>99% lymphocytes); and fraction-IV cells represented the adherent population of fraction-II cells released from the plastic by lidocaine (>95% monocytes). When cells from these fractions were cultured in the presence of erythropoietin, the number of BFU-E-derived colonies was inversely proportional to the number of monocytes present (r = -0.96, P < 0.001). The suppressive effect of monocytes on BFU-E proliferation was confirmed by admixing autologous purified monocytes (fraction-IV cells) with fraction-III cells. Monocyte concentrations of >/=20% completely suppressed BFU-E activity. Reduction in the number of plated BFU-E by monocyte dilution could not account for these findings: a 15% reduction in the number of fraction-III cells plated resulted in only a 15% reduction in colony formation. These results indicate that monocyte-macrophages may play a significant role in the regulation of erythropoiesis and be involved in the pathogenesis of the hypoproliferative anemias associated with infection and certain neoplasia in which increased monocyte activity and monopoiesis also occur.

Cell Communication↗

Endotoxin-stimulated spleen cells: dissociation between DNA synthesis and IgM production at the cellular level.

LPS stimulation of mouse spleen cells in the presence of Hu resulted in almost total suppression of [3H]thymidine incorporation without affecting the percentage of cells induced to produce IgM. Utilizing a method which permitted the simultaneous measurement of IgM production and [3H]thymidine incorporation in individual cells, it was demonstrated directly that LPS stimulation of IgM production can occur in the absence of DNA synthesis.

Animals↗

Capping of the lymphocyte C3 receptor and temperature-dependent loss of C3 rosettes.

The percentage of mouse spleen cells rosetted by bacteria-antibody-complement complexes (BAC) was found to be temperature dependent. Maximal rosetting was observed when BAC and spleen cells were incubated in ice (0.5 degrees C), whereas less rosetting resulted from 37 degrees C incubations. When rosetted cells were incubated at 37 degrees C, cap formation was found to occur, and indirect evidence suggested that the capped bacteria were subsequently shed from the lymphocyte surface. Neither capping nor shedding occurred at 0.5 degrees C, suggesting that these phenomena were responsible for the decreased percentage of rosetted cells found after incubation at 37 degrees C. An incidental observation indicated that a second factor potentially contributing to the loss of rosetted cells at 37 degrees C was the removal of BAC from the surface of cells by phagocytes.

Animals↗