Effect of gamma interferon and interleukin 10 on phosphoinositol turnover by human neutrophils in vitro.

We evaluated the levels of inositolmono-(IP1), di(IP2), tri-(IP3) and tetraphosphates (IP4) in human neutrophils (N) stimulated with gamma interferon (IFN-gamma) (200 microliters from a pool of cell culture supernatant obtained from 1 x 10(7) PHA-primed peripheral blood mononuclear cells (30-60 min at 37 degrees C, 5% CO2)) in the presence of in the absence of interleukin 10 (IL-10) (10 micrograms/10 microliters). The results, reported as mean +/- SEM cpm, showed that IFN-gamma induced a significant increase only in the IP3 level (N + medium = 1,413 +/- 172 and N + IFN-gamma = 8,875 +/- 832). However, this activation mediated by IFN-gamma was blocked partially in the presence of IL-10 (N + IFN-gamma + IL-10 = 2,430 +/- 239) (P < 0.05). Interleukin 10 alone did not induce significant alterations in the content of IP1 (1,203 +/- 123), IP2 (1,880 +/- 163), IP3 (938 +/- 102) or IP4 (2,403 +/- 345) when compared to the respective controls in the absence of IL-10 (IP1 = 1,625 +/- 132; IP2 = 1,343 +/- 149; P3 = 1,413 +/- 172 and IP4 = 3,281 +/- 234). We also demonstrated the inhibitory effect of IL-10 of chemoluminescence generation by human neutrophils during phagocytosis of opsonized particles (OZ). Chemoluminescence generation was enhanced by IFN-gamma (N = OZ = 42.8 +/- 3.9 and N + OZ + IFN-gamma = 66.5 +/- 4.3) and this effect was reduced by IL-10 (N + OZ + IFN-gamma + IL-10 = 37.6 +/- 5.1). These data suggest that IL-10 modulates the neutrophil response and may be important for the development of new treatments of inflammatory injury.

Effect of gamma interferon and interleukin 10 on phosphoinositol turnover by human neutrophils in vitro

We evaluated the levels of inositolmono-(IP1), di-(IP2), tri- (IP3) and tetraphosphates (IP4) in human neutrophils (N) stimulated with gamma interferon (IFN-gamma) (200 mul from a pool of cell culture supernatant obtained from 1 x 10(7) PHA-primed peripheral blood mononuclear cells (30-60 min at 37 degrees Celsius, 5 per cent CO2) in the presence or in the absence of interleukin 10 (IL-10) (10 mug/10mul). The results, reported as mean + SEM cpm, showed that IFN-gamma induced a significant increase only in the IP3 level (N + medium = 1.413 + 172 and N + IFN-gamma = 8,875 + 832). However, this activation mediated by IFN-gamma was blocked partially in the presence of IL-10 (N + IFN-gamma + IL-10 = 2,430 + 239) (P<0.05). Interleukin 10 alone did not induce significant alterations in the content of IP1 (1,203 + 123), IP2 (1,880 + 163), IP3 (938 + 102) or IP4 (2,403 + 345) when compared to the respective controls in the absence of IL-10 (IP1 = 1,625 + 132; IP2 = 1,343 + 149; IP3 = 1,413 + 172 and IP4 = 3,281 + 234). We also demonstrated the inhibitory effect of IL-10 on chemoluminescence generation by human neutrophils during phagocytosis of opsonized particles (OZ). Chemoluminescence generation was enhanced by IFN-gamma (N + OZ = 42.8 + 3.9 and N + OZ + IFN-gamma = 66.5 + 4.3) and this effect was reduced by IL-10 (N + OZ + IFN-gamma + IL-10 = 37.6 + 5.1). These data suggest that IL-10 modulates the neutrophil response and may be important for the development of new treatments of inflammatory injury.

Immune responses during human schistosomiasis mansoni. XVIII. Immunologic status of pregnant women and their neonates.

The immune status of peripheral blood mononuclear cells (PBMC) and sera of pregnant women infected with the helminth Schistosoma mansoni was studied during pregnancy and the cord blood mononuclear cells (CBMC) and sera from their offspring were studied at parturition. PBMC pokeweed mitogen-induced responses were maintained in gravid women, but the responses to both schistosome and non-schistosome antigenic preparations declined progressively during pregnancy. Schistosomal antigens stimulated proliferative responses by the CBMC of many neonates born of infected mothers, but not those of uninfected mothers. These specific responses by CBMC of only neonates born of infected mothers are indicative of in utero, cell-mediated sensitization of the neonates, which could be due either to circulating schistosomal antigens or to anti-idiotypic antibodies which cross the placenta during gestation. Sera from infected mothers and the cord blood sera from their babies showed the same levels of specific IgG anti-schistosomal activity. Anti-schistosomal IgM levels were maintained during pregnancy to some antigens and not to others, while such antibodies were rarely found in cord blood sera, and then only at very low levels.

Idiotypic sensitization in utero of children born to mothers with schistosomiasis or Chagas' disease.

Cord blood mononuclear cells (CBMC) of neonates born of mothers with Chagas' disease or schistosomiasis exhibited strong proliferative responses against idiotypes expressed on antibodies with specificity for Trypanosoma cruzi or Schistosoma mansoni antigens, respectively. These immunoaffinity-purified preparations were stimulatory if they were prepared from pools of patients' sera or from the mother's own serum, taken early during her pregnancy. These CBMC did not respond to normal immunoglobulin, and CBMC of neonates born of uninfected mothers did not respond to antibodies against either T. cruzi or S. mansoni, or normal immunoglobulin preparations. We propose that in utero exposure of a fetus to some idiotypes expressed on placentally transferred antibodies induces anti-Id T lymphocyte sensitization, which we detect as a proliferative response by CBMC exposed to immunoaffinity-purified antibodies expressing the relevant idiotypes. This is the first experimental evidence that children born of mothers with chronic infections undergo natural in utero idiotypic manipulations and are born possessing cellular anti-Id reactivity.

Schistosoma mansoni: cell-mediated immunity evaluated by antigen-induced leukocyte adherence inhibition assay.

Peripheral blood leukocytes (PBL) from schistosomiasis patients fail to adhere to glass in the presence of soluble egg antigens (SEA) or soluble worm adult antigenic preparation (SWAP). These leukocytes are non-reactive with S. mansoni unrelated antigens (C. albicans and bovine albumin). Supernatants obtained from cultures of mononuclear cells of patients with antigens were able to inhibit granulocyte adherence to glass. The inhibition of antigen-induced adherence (LAI assay) was not observed when PBL or supernatants were obtained from normal subjects or from schistosomiasis patients after chemotherapy. These results show that under the conditions tested, leukocytes appear to react directly with SEA or SWAP thus losing their property of adherence to glass.

Schistosoma mansoni: comparison of the killing effect of granulocytes and complement with or without antibody on fresh and cultured schistosomula in vitro.

The efficiency of human granulocytes from patients infected with Schistosoma mansoni in killing either 2-hour or 24-hour schistosomula in the presence of complement was 43.0 +/- 5.0% and 22.7 +/- 1.8%, respectively. Granulocyte, antibody, and complement killed 61.1 +/- 1.7% of 2-hour schistosomula and 13.9 +/- 1.9% of schistosomula previously incubated for 24 hours in complex medium. The resistance of 24-hour schistosomula was less marked when they were incubated in defined medium (20.0 +/- 1.5%). When the 24-hour incubation was performed in the presence of puromycin, however, the death of schistosomula remained high: 51.9 +/- 5.7% with cells plus complement or 48.7 +/- 4.8% with cells, antibody, and complement. The increase in eosinophil percentage in granulocyte suspensions did not enhance their ability to kill 24-hour schistosomula, but it was associated with a slight increase in the degree of damage inflicted on the 2-hour schistosomula.