Antigen presentation is a function of all B cell subpopulations separated on the basis of size

Purified splenic B cells from nonimmune mice were separated by counterflow centrifugal elutriation into 6 subpopulations containing cells of discrete sizes ranging from 119 to 200 μm³. B cells of each subpopulation were competent to process and present a native globular protein antigen, cytochrome c, to a cytochrome c-specific T cell hybrid. In all cases, the B cells' antigen-presenting function was radiation sensitive and did not require T cells or T cell products, since B cells fixed with paraformaldehyde effectively presented a carboxyl-terminal peptide fragment of cytochrome c containing the T cell determinant. Furthermore, the antigen-presenting function of B cells of each subpopulation was augmented by treatment with submitogenic doses of the F(ab')₂ fragment of rabbit anti-mouse Ig antibodies, in that 10-30-fold fewer B cells were required and higher maximal T cell responses were achieved, indicating that B cells of all sizes are capable of being regulated in their antigen presentation function through their surface Ig. In addition, B cells of each subpopulation responded to soluble factors present in the supernatants of activated T cells as evidenced by an increase in volume and by the uptake of [³H]thymidine. These results indicate that B cells, regardless of size, are able to participate in at least two essential phases of T cell-dependent antibody responses, initiating the interaction by processing and presenting antigen to helper T cells and responding to soluble helper factors secreted by activated T cells.     

Compared colonizing and immunizing efficiency of toxinogenic (A+ B+) Vibrio cholerae and an A- B+ mutant (Texas Star-SR) studied in adult rabbits

Four strains of Vibrio cholerae O1 were compared for their ability to colonize and immunize adult rabbit intestine. Three were virulent, toxinogenic (A+ B+) isolates, and one, an A- B+ mutant (Texas Star-SR), was derived by mutagenesis with nitrosoguanidine. When given orally to nonimmune rabbits, virulent strains colonized the small bowel with similar efficiency, whereas Texas Star-SR colonized poorly. Rabbits fed less than 50 CFU of an A+ B+ strain developed marked serotype-specific resistance to recolonization. In contrast, Texas Star-SR evoked resistance to reinfection less efficiently, with a minimum immunizing dose of 10(5) CFU when given once or 10(3) CFU when given twice. Oral inoculation with an A+ B+ strain also evoked vigorous, dose-dependent mucosal antitoxin responses; comparable inocula of Texas Star-SR were much less effective, causing antitoxin responses that were 90 to 95% smaller. Finally, rabbits inoculated once with 10(4) CFU of an A+ B+ strain were markedly protected against experimental cholera or fecal shedding of V. cholerae when challenged with 10,000 times the 50% effective dose of a virulent strain by the RITARD technique. In contrast, an inoculum of 10(4) CFU of Texas Star-SR was nonprotective, and 10(10) CFU was only partially protective. These studies reveal the remarkable efficiency with which virulent V. cholerae evokes intestinal immunity to recolonization or experimental cholera and show that the A- B+ mutant, Texas Star-SR, is substantially less effective.     

Platelet-derived growth factor-BB and transforming growth factor beta 1 selectively modulate glycosaminoglycans, collagen, and myofibroblasts in excisional wounds.

Recombinant platelet-derived growth factor (PDGF) and transforming growth factor beta 1 (TGF-beta 1) influence the rate of extracellular matrix formed in treated incisional wounds. Because incisional healing processes are difficult to quantify, a full-thickness excisional wound model in the rabbit ear was developed to permit detailed analyses of growth-factor-mediated tissue repair. In the present studies, quantitative and qualitative differences in acute inflammatory cell influx, glycosaminoglycan (GAG) deposition, collagen formation, and myofibroblast generation in PDGF-BB (BB homodimer)- and TGF-beta 1-treated wounds were detected when analyzed histochemically and ultrastructurally. Although both growth factors significantly augmented extracellular matrix formation and healing in 10-day wounds compared with controls (P less than 0.002). PDGF-BB markedly increased macrophage influx and GAG deposition, whereas TGF-beta 1 selectively induced significantly more mature collagen bundles at the leading edge of new granulation tissue (P = 0.007). Transforming growth factor-beta 1-treated wound fibroblasts demonstrated active collagen fibrillogenesis and accretion of subfibrils at the ultrastructural level. Myofibroblasts, phenotypically modified fibroblasts considered responsible for wound contraction, were observed in control, but were absent in early growth-factor-treated granulating wounds. These results provide important insights into the mechanisms of soft tissue repair and indicate that 1) PDGF-BB induces an inflammatory response and provisional matrix synthesis within wounds that is qualitatively similar but quantitatively increased compared with normal wounds; 2) TGF-beta 1 preferentially triggers synthesis and more rapid maturation of collagen within early wounds; and 3) both growth factors inhibit the differentiation of fibroblasts into myofibroblasts, perhaps because wound contraction is not required, due to increased extracellular matrix synthesis.     

Animal model of neutropenia suitable for the study of dual-phagocyte systems.

When two sets of phagocytic cells participate simultaneously in the inflammatory process and bacterial killing, the relative contribution of each cell type is difficult to ascertain. The use of cell-specific antibody will permit selective depletion of one phagocyte population. We describe an experimental model of granulocytopenia which utilizes the immunoglobulin G fraction of an antigranulocyte serum. This material markedly depleted circulating polymorphonuclear leukocytes (PMN); within 2 h after injection of antigranulocyte globulin, PMN counts were at 19% of original levels and remained significantly depressed for 24 h. Granulocyte recruitment was also impaired, with only 5 x 10(3) PMN appearing in the lungs in response to an aerosol of Klebsiella, compared to 4.17 x 10(5) PMN in control animals (P less than 0.01). Most importantly, alveolar macrophages retained normal viability (97% versus 94% for control value, P not significant) normal phagocytic function, and normal bactericidal capacity. Antigranulocyte globulin is thus a valuable tool for the study of bacterial defense mechanisms.     

Quantitative displacement of blood acid base equilibrium in acutely water depleted dogs

Summary Dogs were water depleted 2% of their body weight by the intraperitoneal injection of a hyperosmotic solution of saline and glucose (550 mOsm/l). During a 3-h experiment these water-depleted (WD) animals showed a significant decrease in blood pH, base excess (BE) and plasma bicarbonate (Bic_p) and an increase in both hematocrit (Ht) and blood buffer capacity (BBC). These changes were quantitatively time-dependent. The rate change of pH, BE, and Bic_p were –0.072 units h^–1, –4.7 mEq/l h^–1, and 2.9 mEq/l h^–1, respectively. As control dogs showed no significant time-dependent changes in their blood acid-base status, the observed modifications in the experimental dogs were ascribed to water depletion. Increased endogenous acid production related to tissue hypoxia is suggested to be the mechanism that could explain the increased fixed acid accumulation observed in the water-depleted animals.     

Bilateral pneumothoraces secondary to colonic endoscopy

Fiberoptic colonoscopy has significantly altered the managment of patients with colonic diseases. Complications of colonoscopy are not common, but most often include hemorrhage and perforation. This report describes an unusual and interesting case of colonic perforation related to fiberoptic colonoscopy in which the perforation presented as bilateral pneumothoraces.     

Paramagnetic macrocyclic complexes as contrast agents for MR imaging: proton nuclear relaxation rate enhancement in aqueous solution and in rat tissues

Paramagnetic macrocyclic chelates show promise as magnetic resonance (MR) imaging contrast agents due to stability and relaxivity comparable to those of DTPA-type chelates. For the three copper and manganese macrocyclic complexes studied in aqueous solution, T1 and T2 relaxivities ranged from 0.14 to 5.88 mM-1sec-1 at 6.25 MHz. In rats, the intravenous administration of 16 mumol/kg of Mn(cyclam) caused the liver T1 relaxation rate to double at 15 minutes after injection. T1 measurements by pulsed MR imaging and manganese analyses on excised tissue showed that both relaxation rate (1/T1) and manganese content of liver and kidney increase linearly with the dosage of Mn(cyclam). The linear relationship between 1/T1 and manganese content can be considered an "in tissue" relaxivity plot for the agent. The resulting relaxivity is 54 mM-1sec-1 in liver, compared with 3.1 mM-1sec-1 in aqueous solution. Although this work is preliminary, the implication for medical MR imaging applications is that macrocyclic contrast agents can be effective at approximately one-tenth the current typical dose used for gadolinium DTPA.