Interleukin 2 administered in vivo induces the growth of cultured T cells in vivo

The capacity of exogenous IL 2 to induce the growth of antigen-activated T lymphocytes in vivo was evaluated. The in vivo growth of adoptively transferred T lymphocytes that had been previously cultured long-term with IL 2 was initially examined, because in vitro such T cells are exquisitely dependent upon exogenous IL 2 for proliferation and survival. Daily administration of IL 2 in vivo, beginning on the day of cell transfer, induced these IL 2-dependent long-term cultured T lymphocytes to proliferate in vivo, and the magnitude of in vivo growth was proportional to the dose of IL 2 administered. The capacity of IL 2 to induce the in vivo growth of antigen-activated T cells not previously exposed in vitro to exogenous IL 2 was similarly studied. T lymphocytes from the spleens of immune mice, activated by 5-day culture with tumor antigen before transfer, survived poorly in vivo when injected with antigen alone, but demonstrated marked proliferation in vivo in response to antigen and exogenous IL 2. By contrast, immune spleen cells transferred with antigen, but without prior culture, proliferated without supplementary exogenous IL 2. Moreover, the growth of noncultured donor T cells was not augmented by the administration of exogenous IL 2, implying that noncultured spleen cells immune to tumor antigens can produce sufficient amounts of endogenous IL 2 in vivo to sustain maximal T cell growth over the time period examined. Importantly, the ability of exogenous IL 2 to induce donor T cell growth in vivo correlated with its ability to function in vivo to augment the anti-tumor efficacy of specifically immune donor T cells in models for the adoptive therapy of disseminated antigenic murine leukemia. Thus, the current studies highlight the potential of exogenous IL 2 to induce T cell growth in vivo and suggest that the administration of IL 2 in vivo may be useful for augmenting T cell responses that are relatively deficient in the production of endogenous IL 2.     

Enrichment and characterization of clonogenic epithelial cells from adult rat liver and initiation of epithelial cell strains

Summary A highly efficient method is described for obtaining prolifertive epithelial cells from adult rat livers for the reproducible establishment of liver epithelial cell strains. When cells were isolated from livers of 10-to 15-wk-old male Fischer 344 rats by a collagenase-perfusion method, collected by centrifugation at 50×g for 5 min, and cultured in Williams' medium E containing fetal bovine serum and dexamethasone, colonies of epithelial cells different in size and morphology from hepatocytes were obtained. Sequential perfusion with collagenase and dispase yielded numerous epithelial cell colonies. When isolated cells were fractionated by differential centrifugation, the great majority of hepatocytes were sedimented at 50 ×g for 1 min, whereas many non-hepatocytic cells remiined in the supernatant and could be sedimented by a second centrifugation at 50×g for 5 min. Culture of the two fractions revealed that almost all the epithelial cell colonies were derived from cells in the non-hepatocytic cell fraction. The epithelial cells were cytochemically negative for γ-glutamyl transpeptidase activity, whereas an increase in the activity was detected in hepatocytes with duration in culture. Ultrastructural characteristics of hepatocytes were not found in the cells of newly established cell strains. These results suggest that adult rat liver epithelial cells propagable in culture were derived from a cell type other than the hepatocyte.     

Kinetics of microcolony formation of a soil oligotrophic bacterium, Agromonas sp.

Abstract The kinetics of budding/dividing of parent cells at different culture ages, spread on a fresh medium, was formulated by the following model N _t= N _∞ [1 − exp (− λ ( t − t _r)] where N _t is the number of budding/dividing cells in the parent population at time t , N _∞ is the expected number of budding/dividing cells at infinite time, λ is the rate of budding/dividing of parent cells, and t _r is the retardation time. The rate of budding/dividing λ decreased with the increase in the culture age of the parent cell population.     

Dose-response comparisons of canine plasma gastroenteropancreatic hormone responses to bombesin and the porcine gastrin-releasing peptide (GRP)

This study compares the potencies of the porcine gastrin-releasing peptide (pGRP) and bombesin, in causing elevations of canine plasma gastroenteropancreatic (GEP) levels. In the dose range 0-600 pmol . kg-1 . h-1, infusion of both peptides resulted in obvious dose-related elevations of plasma levels of gastrin, pancreatic polypeptide, enteroglucagon, immunoreactive pancreatic glucagon, and insulin. In this dose range, no significant difference in potency between the two peptides in elevating plasma levels of the above hormones was observed. The results of this study, demonstrating equimolar potency of pGRP and bombesin, are in contrast to previous studies reporting that pGRP was less potent than bombesin in causing certain bioactivities in the rat following intracranial administration of the two peptides.     

Mutants of Group D1Salmonella Carrying the Somatic Antigen of Group A Organisms: Isolation and Serological Characterization

O antigen mutants were obtained from Salmonella durban, a group D(1) organism, by treatment with N-methyl-N'-nitro-N-nitrosoguanidine. Serological studies demonstrated that the mutants lost the O-9 antigen factor of the parent organism but acquired the O-2 factor specific to group A Salmonella. Lipopolysaccharides of the mutant strains contained paratose which determines the specificity of O-2 factor. Tyvelose, present in the wild-type lipopolysaccharide, was not found in the mutants. H antigens and other biological characteristics of the mutant strains were the same as those of the wild-type organism. The present finding implies that group A Salmonella species might be derived from group D(1) organisms.     

Mutants of Group D1Salmonella Carrying the Somatic Antigen of Group A Organisms: Evidence for the Lack of Cytidine Diphosphate Paratose-2-Epimerase Activity

The mutant strains of Salmonella durban that possessed O antigen 2, 12 of group A Salmonella were defective in the cytidine diphosphate paratose-2-epimerase activity. The enzyme preparation of the mutant strains catalyzed the conversion of cytidine diphosphate glucose into cytidine diphosphate paratose but not into cytidine diphosphate tyvelose. The defect in the epimerase activity was also confirmed by the use of purified cytidine diphosphate paratose as a substrate. The specificity of dideoxyhexosyl transferase catalyzing the formation of the group-specific determinant is discussed.     

Serodiagnosis of cancer,using porcine antigens recognized by human monoclonal antibody,HB4C5

Antigens, recognized by human monoclonal antibody (HB4C5) generated from a lung cancer patient, were found to occur in porcine pancreas. The antigens-I and -I1 were purified from crude trypsin of porcine pancreas, only by Mono Q column chromatography, and were eluted at 260 and 300 mM NaCl in 10 mM Tris-HCI buffer, pH 7.4, respectively. These antigens differed from trypsin in molecular weight, elution pattern from the Mono Q column, and their reactivity with HB4C5. The molecular weights of the two antigens were almost the same at around 35000. These were used for serodiagnosis with an assay system based on 96-well immunoplates. The reactivities of antigens-I and -II with various sera were similar. When the reactivity of IgG in serum with antigen-II was measured, absorbance at 415 nm in the case of normal and lung cancer patients was 0.178 ± 0.056 and 0.492 ± 0.136 (p < 0.005). The rates of positive reaction in ovary, larynx, uterus, lung and liver cancers were more than 50%, but the rates in stomach and breast cancers were less than 30%. Positive reaction was hardly detected in pancreas cancer and normal controls.     

Initiation and solicitation in male-female grooming in a wild Japanese macaque troop on yakushima island

Grooming initiation among adult males and females of a Japanese macaque troop was analyzed during the non-mating season. Some gestures (“solicitation”) elicited grooming from partners at a high rate. Grooming initiation patterns were divided into two main types: (1) a male often solicited a female to groom him immediately after approaching her and was groomed by her; and (2) a female approached an alpha male selectively, and immediately groomed him. After a female groomed a male, she rarely solicited him to groom her and instead often moved away from him. These results indicated that males were motivated to be groomed, while females were more highly motivated to groom. Sex differences in grooming motivation can be explained by sex differences in the benefit to be groomed.     

Is helodermin-like immunoreactivity in human thyroid C cells due to a salmon calcitonin-like substance?

Helodermin-like and salmon calcitonin (sCT)-like immunoreactivities co-existed in a subset of human calcitonin (hCT)-containing cells in normal human thyroid tissue and medullary thyroid carcinomas. Helodermin/sCT-immunoreactive cells were mostly different from calcitonin gene-related peptide (CGRP)-positive cells. Helodermin and sCT immunoreactivities were not identified in pulmonary and pancreatic hCT-positive neuroendocrine tumors, except for a few lung tumor cells showing positive staining with one of two sCT antisera used. Helodermin immunoreactivity demonstrated by rabbit antiserum R0086 was completely abolished in the presence of synthetic sCT, while sCT immunoreactivity was not absorbed by synthetic helodermin. The carboxyl terminal Arg30-Thr31 sequence (and Pro35 amide structure) of helodermin would be the epitopic site recognized by this antiserum, since a similar amino acid sequence is present in sCT molecules but absent from hCT and CGRP.