A schistosome-expressed immunomodulatory glycoconjugate expands peritoneal Gr1(+) macrophages that suppress naive CD4(+) T cell proliferation via an IFN-gamma and nitric oxide-dependent mechanism.

Lacto-N-fucopentaose III (LNFPIII) is found in human milk and on the Th2 driving helminth parasite Schistosoma mansoni. This pentasaccharide drives Th2-type responses in vivo and in vitro when conjugated to a carrier. In an attempt to further understand early events in Th1 to Th2 switching, we examined phenotypic and functional changes in peritoneal cell populations in BALB/c and SCID mice following LNFPIII-dextran injection. We found that i.p. injection with LNFPIII-dextran resulted in rapid (<20 h) expansion of the Gr1(+) subpopulation of F4/80(+)/CD11b(+) peritoneal cells, comprising up to 75% of F4/80(+)/CD11b(+) peritoneal cells compared with 18% in uninjected or dextran-injected mice. Functionally, these cells suppressed anti-CD3- and anti-CD28-induced proliferation of naive CD4(+) T cells. LNFPIII-dextran also expanded functional Gr1(+) suppressor macrophages in SCID mice, demonstrating that expansion and function of suppressor cells did not require T cells. Suppression in both BALB/c and SCID mice was NO and IFN-gamma dependent, as addition of inhibitors of inducible NO synthase (N(G)-monomethyl-L-arginine), as well as anti-IFN-gamma Abs, restored the ability of CD4(+) T cells to proliferate in vitro. Depletion of the F4/80(+) subset of Gr1(+) cells eliminated the suppressive activity of peritoneal exudate cells showing that these cells were macrophages. Thus, LNFPIII-dextran rapidly expands the Gr1(+) suppressor macrophage population in the peritoneal cavities of otherwise naive mice. These Gr1(+) cells suppress proliferation of naive CD4(+) T cells in an NO-dependent mechanism, and may play a regulatory role in the switching of Th1- to Th2-type responses.

Beryllium, an adjuvant that promotes gamma interferon production.

Beryllium is associated with a human pulmonary granulomatosis characterized by an accumulation of CD4(+) T cells in the lungs and a heightened specific lymphocyte proliferative response to beryllium (Be) with gamma interferon (IFN-gamma) release (i.e., a T helper 1 [Th1] response). While an animal model of Be sensitization is not currently available, Be has exhibited adjuvant effects in animals. The effects of Be on BALB/c mice immunized with soluble leishmanial antigens (SLA) were investigated to determine if Be had adjuvant activity for IFN-gamma production, an indicator of the Th1 response. In this strain of Leishmania-susceptible BALB/c mice, a Th2 response is normally observed after in vivo SLA sensitization and in vitro restimulation with SLA. If interleukin-12 (IL-12) is given during in vivo sensitization with SLA, markedly increased IFN-gamma production and decreased IL-4 production are detected. We show here that when beryllium sulfate (BeSO(4)) was added during in vivo sensitization of BALB/c mice with SLA and IL-12, significantly increased IFN-gamma production and decreased IL-4 production from lymph node and spleen cells were detected upon in vitro SLA restimulation. No specific responses were observed to Be alone. Lymph node and spleen cells from all mice proliferated strongly and comparably upon in vitro restimulation with SLA and with SLA plus Be; no differences were noted among groups of mice that received different immunization regimens. In vivo, when Be was added to SLA and IL-12 for sensitization of BALB/c mice, more effective control of Leishmania infection was achieved. This finding has implications for understanding not only the development of granulomatous reactions but also the potential for developing Be as a vaccine adjuvant.

[The immunomodulating and immunoprotective activity of small ribonucleoprotein complexes (alpha RNPs) from the LRec-1sf cell line]. / Immunomoduliruiushchaia i immunoprotektivnaia aktivnost' malykh ribonukleoproteinovykh kompleksov (al'fa RNP) iz kletok linii LRec-1sf.

Effect of nuclear and released into culture medium alpha RNPs (N- and R-alpha-RNPs, resp.) produced by transformed rat embryo fibroblasts of serum-free cell line LRec-1sf on the nonsensibilized mouse splenocyte cytotoxicity (NK-mediated cell lysis) was studied. A preliminary treatment with N-alpha-RNPs resulted in decreasing K562 cell sensitivity to splenocyte cytotoxicity, whereas pretreatment of the splenocytes themselves exerted no cytotoxic effect. The target cell preincubation with R-alpha-RNPs had no influence on K562 cell resistance to NK cell cytotoxicity. The identical splenocyte preincubation was without action on their cytotoxic effect to LRec-1sf cells, however, resulted in an increase of the K562 cell lysis. The addition of R-alpha-RNPs into splenocyte/target cell mixtures had no influence on NK-mediated lysis, when K562 cells were used as a target cell line, but suppressed the NK-mediated lysis of LRec-1sf cells. The results of the present experiments suggest that alpha RNPs produced by LRec-1sf cell line exhibit the capacity for modulating both mouse NK cytotoxicity, and the transformed cell sensitivity to NK-mediated lysis.

[The natural killer activity of human peripheral blood studied by a radioimmune method and by using flow cytofluorometry]. / Izuchenie aktivnosti estestvennykh killerov perifericheskoi krovi cheloveka radioimmunnym metodom i s pomoshch'iu protochnoi tsitofluorimetrii.

Using flow cytometry and radioimmunological test, a study was made of cytotoxic activities of natural killer (NK) cells of human peripheral blood of three patient groups: healthy donors, persons with infection pathology, and persons being in extreme situations (hyperoxygen gas mixture). With the increase in effector and target (K562) cell amount ratio, NK activity was seen elevating in all groups of patients examined. A correlation was obtained between NK activity in healthy donors and in patients examined by both the methods. NK activity in sick patients, registered by the two methods, was considerably lower than in healthy donors. NK activity in persons that appeared in extreme situations, registered by the radioimmunological test only, did not differ from that in healthy donors, whereas NK activity, registered by the flow cytometry, was found to be considerably lower, resp.

[Monoclonal antibodies to the HNK-1 antigen of human natural killers]. / Monoklonal'nye antitela k HNK1-antigenu estestvennykh killerov cheloveka.

The ability of monoclonal antibodies (MAb) to HNK1 antigen of natural killers (NK), previously purified by affinity chromatography, to react with human blood lymphocytes has been studied by flow cytometry. The results obtained point to some cells, among blood lymphocytes, which have antigenic determinants to these MAbs. A decrease in the NK cell cytotoxic activity was shown after addition of MAb in comparison with the control. This enables us to suggest a reactivity of these MABs to the CD57 leucocyte differential antigens.