scholarly journals Immunophenotyping of blood cells of experimental animals immunized with Brucella abortus thermoextracts

Author(s):  
S. V. Balakhonov ◽  
V. I. Dubrovina ◽  
V. V. Voitkova ◽  
K. M. Korytov ◽  
N. L. Barannikova ◽  
...  

Aim. To study the subpopulational structure of blood cells of the experimental animals immunized with thermoextracts (TE) of Brucella abortus in L- and S-form. Materials and methods. Total 100 certified («Vector», Novosibirsk) outbred mice were immunized with B. abortus I-206 TE in L- and S-form in 20 μg protein dose. After 1, 3, 7, 14 and 21 days of observation the phenotypes (CD45, CD3, CD4, CD8, CD19, CD69) of blood cells were detected. Results. General regularities were revealed after injection of the experimental preparations. So, B. abortus TE in L- and S-form caused the immune response that increased granulocyte number and expression of early activation marker CD69 by T- and B-lymphocytes of blood in early period of observation (1-3 days), decrease in general B-lymphocyte content in late periods of observation (7-21 days). Thus, mice received B. abortus ТE in L-form demonstrated authentically higher CD69 expression of blood lymphocyte subpopulations than mice received B. abortus ТE in S-form. Distinctions in formation of humoral immune response were revealed that probably was connected with alteration of Brucella chemical composition in the course of L-transformation. Conclusion. The investigation established that B. abortus TE in L- or S-form caused immunological reorganization in the experimental animal organisms. On the basis of the fin

Author(s):  
S. V. Balakhonov ◽  
V. V. Voitkova ◽  
V. I. Dubrovina ◽  
K. M. Korytov ◽  
A. V. Korneva ◽  
...  

Aim. To study the subpopulational structure of blood cells of the experimental animals immunized with cellular walls (CW) of Francisella tularensis different subspecies. Materials and methods. Total 210 certified («Vector», Novosibirsk) outbred mice were immunized with CW preparations of F. tularensis different subspecies in 95 pg protein dose. After 3, 7, 10, 14 and 21 days of observation the phenotypes (CD45, CD3, CD4, CD8, CD19, CD69) of blood cells were detected. Results. Dynamics of leukocyte balance in experimental animals depended on F. tularensis subspecies. All studied preparations promoted development of cellular and humoral immunity that confirmed by the increase of monocytes and B-lymphocyte, and also by the expression of CD69 activation marker. CW of F. tularensis subsp. mediasiatica А-61 promoted higher activation of cellular immunity in experimental animals in comparison with other preparations. Probably, exactly this factor causes its high immunogenic activity (83 % survival rate). Conclusion. On the basis of the results there is a necessity to further detailed research of CW immunogenic properties of F. tularensis subsp. mediasiatica А-61 as a perspective component in design of a tularemia vaccine.


2000 ◽  
Vol 192 (7) ◽  
pp. 953-964 ◽  
Author(s):  
Richard K.G. Do ◽  
Eunice Hatada ◽  
Hayyoung Lee ◽  
Michelle R. Tourigny ◽  
David Hilbert ◽  
...  

B lymphocyte stimulator (BLyS) is a newly identified monocyte-specific TNF family cytokine. It has been implicated in the development of autoimmunity, and functions as a potent costimulator with antiimmunoglobulin M in B cell proliferation in vitro. Here we demonstrate that BLyS prominently enhances the humoral responses to both T cell–independent and T cell–dependent antigens, primarily by attenuation of apoptosis as evidenced by the prolonged survival of antigen-activated B cells in vivo and in vitro. BLyS acts on primary splenic B cells autonomously, and directly cooperates with CD40 ligand (CD40L) in B cell activation in vitro by protecting replicating B cells from apoptosis. Moreover, although BLyS alone cannot activate the cell cycle, it is sufficient to prolong the survival of naive resting B cells in vitro. Attenuation of apoptosis by BLyS correlates with changes in the ratios between Bcl-2 family proteins in favor of cell survival, predominantly by reducing the proapoptotic Bak and increasing its prosurvival partners, Bcl-2 and Bcl-xL. In either resting or CD40L-activated B cells, the NF-κB transcription factors RelB and p50 are specifically activated, suggesting that they may mediate BLyS signals for B cell survival. Together, these results provide direct evidence for BLyS enhancement of both T cell–independent and T cell–dependent humoral immune responses, and imply a role for BLyS in the conservation of the B cell repertoire. The ability of BLyS to increase B cell survival indiscriminately, at either a resting or activated state, and to cooperate with CD40L, further suggests that attenuation of apoptosis underlies BLyS enhancement of polyclonal autoimmunity as well as the physiologic humoral immune response.


1991 ◽  
Vol 173 (1) ◽  
pp. 55-64 ◽  
Author(s):  
A K Matsumoto ◽  
J Kopicky-Burd ◽  
R H Carter ◽  
D A Tuveson ◽  
T F Tedder ◽  
...  

The complement system augments the humoral immune response, possibly by a mechanism that involves the B lymphocyte membrane receptor, CR2, which binds the C3dg fragment of C3 and triggers several B cell responses in vitro. The present study demonstrates that CR2 associates with a complex of membrane proteins that may mediate signal transduction by ligated CR2. Monoclonal antibodies to CR2 immunoprecipitated from digitonin lysates of Raji B lymphoblastoid cells a membrane complex containing CR2, approximately equimolar amounts of CD19, which is a member of the immunoglobulin superfamily, and three unidentified components: p130, p50, and p20. The complex, which was immunoprecipitated also with anti-CD19, could be dissociated by Nonidet P-40, accounting for its absence in previous studies of CR2. Expression of recombinant CR2 and CD19 in K562 erythroleukemia cells led to formation of a complex that contained not only these two proteins but also p130, p50, and p20, and another component, p14. These unidentified components of the CR2/CD19 complex coimmunoprecipitated with CD19 and not with CR2 from singly transfected cells, indicating primary association with the former. CD19 replicated the capacity of CR2 to interact synergistically with mIgM for increasing free intracellular Ca2+, suggesting that the complex mediates this function of CR2. Therefore, CR2 associates directly with CD19 to become a ligand-binding subunit of a pre-existing signal transduction complex of the B cell that may be representative of a family of membrane protein complexes. This interaction between the complement and immune systems differs from that between immunoglobulin and Clq by involving membrane rather than plasma proteins, and by having complement involved in the afferent phase of the immune response.


1998 ◽  
Vol 66 (4) ◽  
pp. 1764-1767 ◽  
Author(s):  
Odile Poulain-Godefroy ◽  
Nathalie Mielcarek ◽  
Nathalie Ivanoff ◽  
Franck Remoué ◽  
Anne-Marie Schacht ◽  
...  

ABSTRACT In an attempt to increase the immunogenicity of mucosally delivered antigens, we incorporated the Bordetella pertussisfilamentous hemagglutinin (FHA) adhesin into liposomes containing the glutathione S-transferase of Schistosoma mansoni (Sm28GST) as a model antigen. Outbred mice immunized twice intranasally with liposomes containing a constant suboptimal dose of Sm28GST and increasing doses of FHA produced anti-Sm28GST antibodies in a FHA dose-dependent manner. The addition of 3 μg of FHA to the liposomes induced more than 10-fold-higher anti-Sm28GST antibody titers, compared to those induced by liposomes without FHA. The presence of FHA did not alter the nature of the humoral immune response, and the sera contained anti-Sm28GST immunoglobulin G1 (IgG1), IgG2a, and IgG2b. However, anti-Sm28GST IgA was only detected when at least 3 μg of FHA was added to the preparation. These results show a promising potential for FHA to enhance the immunogenicity of mucosally administered antigens incorporated into liposomes.


Author(s):  
Losa Rose ◽  
Bablu Kumar ◽  
Ankita Jain ◽  
M K Singh ◽  
Abhishek .

Outer membrane vesicles (OMVs) contain biologically active proteins, lipoolysaccharide (LPS), periplasmic and membrane-bound proteins and are known to perform diverse biological functions. OMVs from Brucella abortus S19 were isolated and characterized by transmission electron microscopy (TEM), SDS-PAGE and immunoreactivity was investigated by western blotting. On TEM, bilayered spherical structures of 50-200 nm were observed. SDS-PAGE of OMVs revealed approximate bands size of 82 kDa, 68 kDa, 38 kDa, 32 kDa, 29 kDa and 18 kDa. Western blot analysis of OMVs revealed a dominant immunoreactive band of 38 kDa that correspond to some major outer membrane proteins. Humoral immune response was measured by indirect ELISA which showed that OMV specific antibodies were detected from 7th day post immunization (DPI) onwards and showed a rising trend up to 35th DPI. Cell mediated immune (CMI) response against OMVs as evidenced by the proliferation of splenocytes have also been observed. Thus OMVs were found to possess immunogenic proteins which had potential to induce both humoral as well as cell mediated immunity. After correlating this immune response with protection it has been concluded that OMV can be used as one of the vaccine candidate against brucellosis.


1986 ◽  
Vol 14 (01n02) ◽  
pp. 33-36
Author(s):  
G.M. Sein ◽  
M. Phil

The effects of Korean herbal medicine (B.C.L.) on some parametrs of immunological response were studied in mice, B.C.L. pretreatment given either intraperitoneally subcutaneously in a dose of 0.75 mg/mouse did not significantly inhibit lymphocyte transformation induced by concanavalin A. However, B.C.L. pretreatment in a dose of 2.25 mg/mouse was found to reduce significantly both the plaque-forming cells to sheep red blood cells immunisation as well as total splee cell population. Thus, B.C.L. pretreatment with a higher does (2.25 mg/mouse) can selectively depress the humoral immune response. It is unclear, however, whether this action is mediated by the parent compound or its metabolities.


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