Response Of Mouse Spleen Cells Research Articles

Biochemical characterisation of lectin from wild chickpea (Cicer reticulatum L.) with potential inhibitory action against human cancer cells.

A wild chickpea lectin (WCL) from the seeds of Cicer reticulatum L. was chromato-purified using DEAE-Cellulose and SP-Sephadex ion exchange chromatography. WCL was thermostable upto 60°C with broad pH optima (pH 5-9) and various divalent metal ions did not influence its activity. WCL demonstrated DNA protection in a dose-dependent manner. The lectin exerted antifungal activity against diverse fungal pathogens. WCL augmented the mitogenic response of mouse spleen cells at 10µg/ml concentration and showed an inhibition of HIV-1 reverse transcriptase at IC50 of 200µM. Against human cancer cell lines, lectin demonstrated anticancer potential. The cell viability assay in HepG2, Ishikawa, MCF-7 and MDA-MB-231 cell line demonstrated IC50 values of 61.8, 54.4, 37.5 and 44.2µg/ml respectively. PRACTICAL APPLICATIONS: WCL exhibited distinct medicinal properties vis-à-vis antiproliferative, mitogenic, antifungal/bacterial and HIV-1 reverse transcriptase inhibiting activities. The potential of WCL can be a subject of exploration from a pharmacological standpoint.

In vitro Studies of the Antibody Response: Antibodies of Different Specificity are Made in Different Populations of Cells.

In vitro Studies of the Antibody Response: Antibodies of Different Specificity are Made in Different Populations of Cells.

Enhanced Gamma Interferon Responses of Mouse Spleen Cells following Immunotherapy for Tuberculosis Relapse

Gamma interferon responses of spleen cells in mice were examined during postchemotherapy relapse of intraperitoneally induced latent tuberculous infection. The mycobacterial extract RUTI, which prevented the relapse, significantly enhanced the immune responses to secreted and structural recombinant mycobacterial antigens, suggesting that RUTI-mediated protection was mediated by activated T cells.

In Vitro Immunomodulatory Activity of Bo‐Yang‐Hwan‐O‐Tang

Bo‐yang‐hwan‐o‐tang (BHT), an herbal decoction has been mainly used for improvement of blood flow in oriental medicine. Its in vivo immunomodulation was recently demonstrated but the effective mechanisms have not been described. This study was carried out to evaluate in vitro immunomodulatory activity of BHT. Water extract of BHT significantly promoted in vitro proliferative responses of mouse spleen cells (SPC) and also further enhanced the proliferation of SPC stimulated with anti‐CD3 antibody. Unexpectedly, addition of BHT extract did not affect proliferation of both resting and CD3‐activated T cells, whereas it showed a strong mitogenic activity on B cells. Flow cytometric analysis of CFSE‐stained SPC showed that BHT‐mediated enhancement of CD3‐activated SPC proliferation is due to T cell, but not B cell, division. Mixed culture experiment combining T and mitomycin C‐treated B cells demonstrated that BHT‐mediated enhancement of CD3‐activated T cell proliferation was dependent on the presence of B cells. However, B cell‐derived factors were not involved in BHT effect on T cell proliferation. In the presence of B cells, BHT treatment resulted in a great enhancement in IL‐2 production of CD3‐activated T cells, and BHT effect on T cell proliferation was completely abrogated by addition of exogenous IL‐2, indicating that IL‐2 plays a critical role in BHT‐mediated enhancement of CD3‐activated T cell proliferation. Taken together, our data revealed that BHT possesses a potent B cell mitogenic activity and also can enhance activated T cell response through B cell regulation.

Preparation of casein phosphopeptides from different kinds of pasteurized milks and their action on proliferative responses of mouse spleen cells in cell cultures

Preparation of casein phosphopeptides from different kinds of pasteurized milks and their action on proliferative responses of mouse spleen cells in cell cultures

Isolation and Characterization of a Cytotoxic Pentapeptide, κ-casecidin, from Bovine κ-casein Digested with Bovine Trypsin

Bovine κ-casein produced cytotoxic activity toward mouse spleen cells when it was digested with bovine trypsin or chymotrypsin. The cytotoxic activity reached a maximum when the casein was digested with the trypsin at 45°C for 5h and was observed when the spleen cells were cultured for 6h with the digest. A cytotoxic peptide was isolated from a digest prepared from κ-casein treated with trypsin for 5h by a combination of ion-exchange column chromatography, gel filtration and reversed-phase high performance liquid chromatography. The sequence of the isolated peptide was estimated as Phe-Phe-Ser-Asp-Lys, being compatible with sequence 17-21 of bovine κ-casein. A chemically synthesized peptide, Phe-Phe-Ser-Asp-Lys, produced cytotoxic activity toward mouse spleen cells at a dose response manner. On the other hand, the isolated peptide retained cytotoxic activities toward animal cells such as mouse spleen T- and B-lymphocyte-enriched cells, bovine milk cells and Jurkat, Clone E6-1 cells (a cell line from human acute leukemic T-lymphocyte). The mouse lymphocytes cultured with the isolated peptide revealed an internucleosomal DNA fragmentation. The isolated peptide significantly inhibited the proliferative responses of mouse spleen cells stimulated by lipopolysaccharide, pokeweed mitogen, phytohemagglutinin and concanavalin A. Moreover, the isolated peptide showed notable antibacterial activity against Staphylococcus aureus IFO3060.

Influe of Berberine On T-Cell Mediated Immunity

The protoberberine alkaloid berberine is isolated as a main alkaloid from the roots and bark of Berberis vulgaris. Berberine strongly inhibited in vitro the proliferative response of mouse spleen cells to T-dependent mitogens concanavalin A (Con A) and phytochemagglutinin (PHA). Spleen cells obtained from berberine-treated mice (10 mg/kg/3 days) expressed enhanced proliferative response to both mitogens. Berberine was applied to mice at different intervals before or after the induction of adjuvant arthritis (AIA) and Candida albicans (C. albicans) infection. The application of the alkaloid to new born mice (5 days after birth at a dose of 5 mg/kg/3 days) did not change the course of AIA and C. albicans infection. Its application at three 10 day intervals (5 mg/kg), starting from the 5 day after birth increased the joint inflammation in AIA. The host resistance to C. albicans infection was not affected, while the delayed type hypersensitivity (DTH)-reaction against the pathogen was enhanced. The alkaloid inhibited the development of AIA when applied after its onset (10 mg/kg from day ±3 to ±12 day). Berberine treatment during the ongoing infection did not influence its outcome (from ±2 to ±10 day).

Inhibition of mitogen-induced proliferative responses of lymphocytes by bovine kappa-caseinoglycopeptides having different carbohydrate chains.

Bovine kappa-caseinoglycopeptides (i.e. residues 106-169, CGP) were prepared from kappa-casein digested with rennin and a commercial whey protein concentrate. CGP from whey protein concentrate was further divided into seven CGP fractions having different carbohydrate compositions using FPLC. Unfractionated CGP inhibited lipopolysaccharide (LPS)- and phytohaemagglutinin (PHA)-induced proliferative responses of mouse spleen cells and rabbit Peyer's patch cells. The unfractionated CGP also inhibited antibody responses to sheep red blood cells in mouse spleen cell cultures. However, seven CGP fractions having zero to five N-acetylneuraminic acid (NANA) residues had different inhibitory effects on both LPS- and PHA-induced proliferative responses of mouse spleen cells. The inhibitory effect on PHA-induced proliferative responses increased with increasing numbers of NANA residues, whereas that on LPS-induced proliferation was highest with the CGP fraction having two NANA residues. Both inhibitory effects decreased significantly after neuraminidase or chymotrypsin digestion. These findings indicate that both the carbohydrate (particularly the NANA residues) and the polypeptide portions are essential for inhibitory effects on LPS- and PHA-induced proliferative responses of mouse spleen cells.

Inhibition of concanavalin a‐induced proliferative responses of mouse spleen cells and rabbit Peyer's patch cells by hen egg‐white avidin

Hen egg‐white avidin inhibited the proliferative responses of mouse spleen cells stimulated by both concanavalin A (Con A) and pokeweed mitogen. It also inhibited mitogen‐induced proliferation of rabbit Peyer's patch cells. The Con A‐induced proliferation of spleen cells was inhibited by streptavidin without α‐D‐mannose residues as well as by egg‐white avidin with α‐D‐mannose residues. Egg‐white avidin was found to bind directly to spleen cells, but it had no cytotoxic activity against spleen cells or Peyer's patch cells. Egg‐white avidin did not appear to inhibit the formation of cytokines required for Con A‐induced spleen cell proliferation or to induce the production of growth‐inhibiting cytokines. On the other hand, egg‐white avidin's inhibitory effect on Con A‐induced spleen cell proliferation decreased as the time to addition of egg‐white avidin increased, and was lowest when the protein was added after 10 h of spleen cell cultivation with Con A. Spleen cells cultured with Con A resulted in a signif...

Cellular basis of differential responsiveness of lymph nodes and spleen to 23-valent Pnu-Imune vaccine.

Subcutaneous administration with 23-valent pneumococcal polysaccharide vaccine (Pnu-Imune) elicited a good antipneumococcal capsular polysaccharide response from the spleen but not from peripheral lymph nodes (LN) (cervical, brachial, axillary, popliteal, inguinal) of BALB/c mice. To evaluate the cellular basis of the unresponsiveness of LN to this vaccine and to identify the cell types in spleen that are necessary for induction of polysaccharide Ab responses, a reliable in vitro culture system was developed to obtain plaque-forming cell responses to the Pnu-Imune vaccine. The in vitro antipneumococcal polysaccharide response of mouse spleen cells was found to be relatively T cell independent. Unlike spleen cells, LN cells did not respond to the Pnu-Imune vaccine in vitro. However, LN cells responded to Pnu-Imune vaccine when co-cultured with irradiated spleen cells or splenic dendritic cells but not with accessory cell-depleted spleen cells. LN cells were found to contain the necessary accessory cells that can support the polysaccharide response but in reduced numbers. The LN response to the vaccine was also reconstituted by IL-1 or Th2-associated cytokine IL-5. In contrast, IFN-gamma, a Th1 cell-derived lymphokine was able to suppress the in vitro splenic response to the vaccine suggesting that differential activation of Th1 and Th2 types of Th cells might be one mechanism by which T cells regulate Ab responses to pneumococcal polysaccharides.

Indonesian Medicinal Plants. XI. Chemical Structures of Caesaldekarins a and b, Two New Cassane-Type Furanoditerpenes from the Roots of Caesalpinia major (Fabaceae).

Five new cassane-type furanoditerpenes named caesaldekarins a (1), b (2), c, d, and e were isolated from the roots of Caesalpinia major (Fabaceae), an Indonesian medicinal plant. The chemical structures of 1 and 2 have been elucidated on the bases of physicochemical evidence and chemical derivations. The major diterpenoid caesadekarin a (1) was shown to inhibit mitogen responses of mouse spleen cells and interleukin-1 production.

Species‐Nonspecific Action of Soluble Immunosuppressive Factor Produced by Murine Immature Erythroid Cells

Immature murine erythroid cells have natural suppressor cell activity suppressing in vitro mouse plaque-forming cell response and LPS-stimulated proliferative response of mouse spleen cells. A cell population enriched by erythroid precursors can produce a soluble activity with similar suppressive abilities. This erythroid suppressor activity (ErSF) has a low Mr (1 to 10 kDa). It can suppress both IgM and IgG secretion of PWM-stimulated human B cells and inhibit the proliferative response of PWM-stimulated human B lymphocytes, thereby indicating a species-nonspecific type of action. Antigen-induced mouse PFC response and mitogen-stimulated mouse and human B cell proliferation were inhibited dose dependently by the addition of ErSF, which did not significantly reduce cell viability, however. Kinetic studies showed that ErSF suppressed Ig secretion and proliferation of mitogen-stimulated B lymphocytes in all culture periods. The suppressive effect of ErSF on B cell proliferation was indirect, requiring the presence of IL-2 (but not of IL-1 alpha, IL-1 beta, or BCGF). In the presence of IL-2, the proliferative response of SAC-stimulated B cells was suppressed by ErSF when added at all time intervals studied but was maximal when ErSF was added on the third day of culture. The role of suppressor factor produced by immature murine erythroid cells in the phenomenon of "natural suppression" has been discussed.

Activation of T and B cells by a crude extract of Artocarpus integrifolia is mediated by a lectin distinct from jacalin

The biological activities previously described for a crude extract derived from seeds of Artocarpus integrifolia (jack fruit) are shown in the present work to be assigned to two distinct fractions present in this extract. One fraction is the D-galactose binding lectin, jacalin, obtained by affinity purification on a D-galactose Sepharose column. The other fraction is a D-mannose-binding protein which we propose to call ‘Artocarpin’. As is well documented, jacalin binds to human IgA 1 and is a useful tool for the purification of this immunoglobulin. We show here that the remaining biological activities consisting of the proliferative response of mouse spleen cells and human peripheral blood mononuclear cells and polyclonal activation of human and mouse B cells for the secretion of immunoglobulin are mediated by artocarpin. Artocarpin is unique in its capacity to induce polyclonal activation of B cells in the absence of proliferation. BALB/c nu/nu spleen cells failed to proliferate which indicates that this lectin is a T cell-dependent B cell polyclonal activator.

Cell-mediated immunity in mice infected with Acanthamoeba culbertsoni

Observations were made on the differences of cell-mediated responses in mice of three infection groups differently scheduled in their severity with pathogenic Acanthamoeba culbertsoni. Infections were done by dropping 5 microliters saline suspension containing 3 x 10(3), 1 x 10(4), or 1 x 10(5) trophozoites, respectively. Amoebae were cultured axenically in CGV medium and inoculated into the right nasal cavity of C3H/HeJ mice aging around 6-8 weeks, under the anesthesia by intraperitoneal injection of secobarbital. Delayed type hypersensitivity (DTH) responses in footpad and blastogenic responses of mouse spleen cells using (3H)-thymidine and the serum antibody titer were measured up to day 14 after infection, and natural killer cell activities were measured up to day 5 after infection. The results obtained in this study were as follows: 1. The mice infected with 3 x 10(3) trophozoites showed mortality rate of 17%, and 34% in the mice infected with 1 x 10(4) trophozoites and 65% with 1 x 10(5) trophozoites. 2. In regard to DTH responses in all experimental groups, the level increased on day 7 and declined on day 14 after infection, but their differences could not be noted between infected and control groups. 3. The blastogenic responses of splenocytes treated with amoeba lysates and lipopolysaccharides (LPS) showed no difference from the control group. The blastogenic responses of splenocytes treated with concanavalin A were declined significantly in the experimental group as compared with the control group, but the blastogenic responses of splenocytes treated with polyinosinic acid were not different from the control group. There was also no difference among three infected groups. 4. The cytotoxic activity of the natural killer cells was activated on day 1 after infection and declined to the level of control group on day 2 in all experimental groups. On day 5 after infection, the natural killer cell cytotoxicity was significantly suppressed as compared with the control groups. 5. The serum antibody titers of the infected mice increased after day 7, but there was no statistical difference between the three infected groups. In summary of the results, there was no difference in cell-mediated immune responses of three experimental groups scheduled with different infection intensities. But there was a significant difference in cell-mediated immune responses between infected and control mice. It is considered that cell-mediated immune responses should be involved in murine model infected with A. culbertsoni.

Reduced proliferative response of mouse spleen cells to mitogens during infection with Salmonella typhimurium or Listeria monocytogenes

A significant reduction in the mitogenic responsiveness (uptake of 3H-thymidine) of murine spleen cells to concanavalin A, phytohemagglutinin or lipopolysaccharide was observed during infection with virulent Salmonella typhimurium. The decreased response to mitogens could be observed independent of the immunity to typhimurium ( Ity) genotype, i.e. in CBA/J mice and C 3H/HeJ mice ( Ity r) as well as in C57BL/6 mice ( Ity s). Because reduced responsiveness was demonstrated in C 3H/HeJ mice, which are susceptible to S. typhimurium infection but are unresponsive to lipopolysaccharide, it is concluded that the two phenomena are not correlated with one another. A similar decrease in response to mitogens was shown in mice infected with Listeria monocytogenes. Reduction in mitogenic responsiveness was directly correlated with the number of viable bacteria detected in the spleen cell suspension. Decreased lymphoproliferation could be observed as early as 2 days after infection and lasted 3 weeks in sublethally infected mice. The question remains whether or not the reduced responsiveness indicates an enhanced susceptibility to infection or merely represents a high degree of activation of defense mechanisms.

Suppression of the cytotoxic T-lymphocyte response in mice by pertussis toxin

Pertussis toxin (PT), the major toxin produced by Bordetella pertussis, has been reported both to enhance and to suppress immune responsiveness. These findings suggested that PT contributes to the virulence of B. pertussis through mechanisms involving immune regulation. We report that PT suppressed both the primary and the secondary cytotoxic T-lymphocyte (CTL) responses of mouse spleen cells cultured against two different allogeneic stimulator spleen cells in vitro. This suppression was dependent on the dose of PT used. PT must be present during the initial stages (within the first 24 hr) of CTL generation. Soluble factor (s) obtained from spleen cells preexposed to PT did not suppress the CTL response. Suppression of the CTL response observed was not due to depletion of the antigen by PT. The cytotoxic activity of CTL clones could not be suppressed by PT. The Analysis of responder spleen cells, fractionated by antiimmunoglobulin panning techniques, provided evidence that L3T4 −, Lyt 2 + cells mediate the PT-induced immunosuppression. We propose that suppression of the CTL response by PT is generated through the activation of L3T4 −, Lyt 2 + suppressor T lympohcytes.

Legionella pneumophila-induced suppression of early blastogenic responsiveness of murine spleen cells to specific and nonspecific stimulation in vitro

Legionella pneumophila whole cells, including viable organisms or a killed vaccine, early after injection into mice suppressed the blastogenic responses of mouse spleen cells to both specific (i.e.,Legionella) and nonspecific (i.e., plant mitogen andEschericia coli lipopolysaccharide) stimulators. Mice given injections of sublethal numbers of viableLegionella or of a killed vaccine evidenced 3–4 weeks thereafter a marked increase in blastogenic sensitivity of their spleen cells to theLegionella antigen, either whole cells or soluble antigen, but no increase in responsiveness to nonspecific mitogens (i.e., concanavalin A, phytohemagglutinin, andE. coli lipopolysaccharide) was evident. In contrast, during the first week or so after injection of mice with either viable or killedLegionella, marked suppression of blastogenic responsiveness of spleen cells toLegionella antigens was evident. Concomitant suppression also occurred to concanavalin A and phytohemagglutinin, as well as toE. coli lipopolysaccharide. However, by the second week after injection of the animals with live or killedLegionella, such suppression disappeared. The importance of such early specific suppression of a cellular immune response early after exposure toLegionella antigen, in contrast with the early and sustained rise in specific antibody formation is being further investigated.

Immunological responsiveness of mouse spleen cells after in vivo or in vitro exposure to 3-acetyldeoxynivalenol

The effects of 3-acetyldeoxynivalenol (3-AcDON) on mitogen-induced lymphocyte proliferation and antibody production were studied in male CD-1 mice exposed to 0, 2.5, 5 or 10 ppm 3-AcDON in the diet for 35 days. Mitogen-induced lymphocyte proliferation and T-cell-independent antibody responses to dinitrophenyl-ficoll or Escherichia coli were not altered by dietary exposure to 3-AcDON. The T-cell-dependent antibody response to sheep red blood cells was increased in the group fed 10 ppm 3-AcDON. In vitro, 3-AcDON inhibited lymphocyte proliferation in a dose-dependent manner. Inhibition was observed when the toxin was present during the first 8 hr in phytohaemagglutinin-stimulated cultures and during the first 24 hr in lipopolysaccharide-stimulated cultures. This suggests that 3-AcDON blocks an early step in lymphocyte activation. This inhibition was not restored by thiol reducing agents (dithiothreitol, l-cysteine or 2-mercaptoethanol). Similarly, the addition of lymphokines, including interleukin-1 or interleukin-2, did not alter the inhibitory effects of 3-AcDON. These results suggest that the in vitro effects of 3-AcDON may not reflect its in vivo immunotoxicity. However, 3-AcDON may serve as a chemical probe for examining the activation process of lymphocyte proliferation.

Effects of bifonazole, fluconazole, itraconazole, and terbinafine on the chemiluminescence response of immune cells.

The luminol-enhanced chemiluminescence (CL) assay was used to examine the effects of antifungal agents tested at concentrations above and below therapeutically achievable levels on the CL response of mouse spleen cells. Reduction in the CL response of phagocytic cells may be indicative of an inhibition of the cellular immune response. Concomitantly, an increase in the CL response of phagocytic cells may indicate an enhancement of the immune capacity of these cells. The effects of four antifungal agents, bifonazole, fluconazole (UK-49,858), itraconazole, and terbinafine were studied. Changes in the CL response were assessed in terms of peak intensity, time to peak intensity, and area under the intensity-time curve compared with appropriate diluent controls for each drug. Both bifonazole and itraconazole caused significant reduction in peak CL intensity only at the highest level assayed (20 mg/l). Fluconazole had no significant effect on the CL response of mouse spleen cells at levels up to 20 mg/l, inclusive. Although terbinafine had no significant effect on peak CL intensity, it did cause a significant decrease in time to peak response at levels above 5 mg/l. This decrease in time to peak response may be indicative of an enhancement in the immune capacity of the mouse spleen cells; the clinical significance of this observation remains to be determined.

Influence of six antifungal agents on the chemiluminescence response of mouse spleen cells.

The luminol-enhanced chemiluminescence (CL) assay is a measure of the early events of phagocytosis, leukocyte activation, and immune cell interactions. Reduction in the CL response of immune cells may be indicative of an inhibition of the immune response. This study was undertaken to examine the effects of antifungal agents tested at concentrations above and below therapeutically achievable levels on the CL response of mouse spleen cells. The effects of six antifungal agents--amphotericin B, ketoconazole, miconazole, 5-fluorocytosine, Bay-n-7133, and ICI 153,066--were studied. Changes in the CL response were assessed in terms of peak intensity and area under the intensity-time curve compared with appropriate diluent controls for each drug. Only amphotericin B and ketoconazole caused statistically significant lowering of the peak intensity at levels that are therapeutically attainable (mean peak plasma levels of 2 to 4 and 3.5 to 16 micrograms/ml, respectively). Although amphotericin B and ketoconazole caused reduction in the CL response, removal of the unbound drug in the preparation by centrifugation, washing, and suspension of cells in fresh drug-free medium resulted in a return of CL activity equivalent to the activity observed in cells not exposed to these agents. These results suggest that amphotericin B and ketoconazole at therapeutic concentrations may cause a reduction in immune cell antimicrobial activity; the clinical significance of these observations remains to be determined.