Soluble recombinant enterovirus 71 VP1 fused to truncated newcastle disease virus nucleoprotein elicits immune responses in mice.

We have studied the capability of the mouse thymus for asymmetrical formation. Concanavalin A (Con A)-stimulated proliferation of thymocytes from the right and left lobes of the thymus appeared to be significantly different. The direction of the differences depends on the dominance of the brain hemispheres with regard to motor asymmetry. In mice with right-dominant hemispheres, thymocytes from the left lobe of the thymus demonstrate a higher level of Con A-stimulated proliferation than those from the right lobe. In mice with left-dominant hemispheres, we found the opposite dependence. The in vivo experiments showed that the properties of cells from the contralateral lobes of the thymus proved to be a deciding factor that defines the differences at the level of the immune response in recipient mice with left-dominant hemispheres. This effect was less pronounced in mice with right-dominant hemispheres. Further analysis showed that left and right-dominant hemisphere mice differ according to the immune response only if mice from both groups received cells from the left but not from the right lobe of the thymus. That is, in the formation of the immune response to sheep red blood cells, the functional asymmetry of both the brain and thymus is of great importance. The experiments show that brain hemispheres and cells from different lobes of the thymus are able to interact in the regulatory effect on the immune response. The injection of cells from the thymus lobe ipsilateral to the dominant hemisphere, results in a significant excess of the immune response in left-dominant hemisphere mice in comparison with the response of right-dominant hemisphere mice. It can be concluded that this work demonstrates, for the first time, the asymmetrical function of a bilateral immune organ – the thymus. The asymmetry is shown not only at the level of Con A-induced proliferative activity but also at the level of the influence on the humoral T-dependent immune response in mice. Besides, we have found the interaction of brain hemispheres and thymus lobes in the regulation of the immune response.

Lycium barbarum polysaccharide (LBP) is isolated from the fruit of Chinese herbal Lycium barbarum. Previous studies had demonstrated that LBP could inhibit tumor growth and enhance the immunity in mice. However, the effect of LBP on systemic and local immune responses in vivo, especially on phenotypic and functional changes of T cells, is still largely unknown. In the present study, we investigated the effects of LBP on systemic and local T cell-dependent antitumor immune responses in H22 tumor-bearing mice. The results showed that LBP could inhibit the solid tumor growth in mice, but showed little effect on the body weight or spleen index. Furthermore, LBP could maintain high levels of T cells in peripheral blood (PB), tumor draining lymph node (TDLN), and tumor tissue, prevent the increase of Tregs while promote infiltration of CD8+ T cells in tumor tissue, inhibit the production of TGF-β1 and IL-10 in serum, decrease the exhaustion phenotype of T cells, and maintain cytotoxicity of lymphocytes. Taken together, our results demonstrated that LBP simultaneously induced systemic and local immune responses in H22 tumor-bearing mice by alleviating immunosuppression and maintaining antitumor immune responses in mice.

To study the effect of battery industry effluent on cell-mediated immune response (CMI) in mice, twoweek-old (256) mice were randomly divided into 4 equal groups namely, control (group 1), vaccinated (group 2), effluent treated (group 3) and vaccine+effluent treated (group 4). Groups 2 and 4 mice were immunised with Ranikhet disease vaccine (R2b) at every 15 days up to 120 days. Group I was kept as control while group 3 and 4 mice were given battery industry effluent daily orally ad libitum instead of drinking water. Cellular immune response in mice was measured by the lymphocyte stimulation test (LST), total leucocyte count (TLC) and absolute lymphocyte count (ALC). The results showed a significant decrease in lymphocyte stimulation index, total leukocyte count and absolute lymphocyte count leading to suppression of CMI response in battery industry effluent fed mice in comparison to control.

Homeostasis-driven proliferation of T cells is an important means of reconstituting T-cell-dependent immunity after lymphodepletion regimens, such as chemotherapy or radiotherapy. Immune-reconstituted mice that receive a tumor vaccine mount more efficient anti-tumor immune responses compared with control mice. In the present study, we evaluated the anti-tumor immune responses in immune-reconstituted mice vaccinated with inactivated leukemia cells and explored the mechanisms underlying these immune responses. Test C57BL/6 mice were lymphodepleted by irradiation and immune-reconstituted with naïve mouse spleen lymphocytes. Mice were then injected with an inactivated FBL-3 tumor cell vaccine and challenged with FBL-3 tumor cells. Anti-tumor responses were evaluated by determining the rate of tumor formation, latency, tumor size, interferon gamma levels, and macrophage and CTL cytotoxicities. When challenged with tumor cells, immune-reconstituted, vaccinated mice exhibited a significantly lower mortality, smaller average tumor volume, and a significantly longer mean survival time. They had more robust cellular immunity, reflected by higher levels of INF-γ production and higher macrophage- and CTL-mediated cytotoxicities. Our results suggest that immune reconstitution enhanced the anti-tumor immune responses in mice injected with a tumor vaccine via generation of CTLs. These results have important implications for immunotherapy used for leukemia.

To study the effect of battery industry effluent on cell-mediated immune response (CMI) in mice, twoweek-old (256) mice were randomly divided into 4 equal groups namely, control (group 1), vaccinated (group 2), effluent treated (group 3) and vaccine+effluent treated (group 4). Groups 2 and 4 mice were immunised with Ranikhet disease vaccine (R2b) at every 15 days up to 120 days. Group I was kept as control while group 3 and 4 mice were given battery industry effluent daily orally ad libitum instead of drinking water. Cellular immune response in mice was measured by the lymphocyte stimulation test (LST), total leucocyte count (TLC) and absolute lymphocyte count (ALC). The results showed a significant decrease in lymphocyte stimulation index, total leukocyte count and absolute lymphocyte count leading to suppression of CMI response in battery industry effluent fed mice in comparison to control.

Depression of the cellular immune responses in mice with disseminated histoplasmosis is associated with deficient production of interleukin-2 (IL-2) by splenocytes. Therefore, we examined whether a highly purified preparation of IL-2, recombinant human IL-2 (rIL-2), could modify the cellular immune responses in infected mice and whether this lymphokine could alter the severity of histoplasmosis in animals. Exogenous rIL-2, at concentrations of up to 1,000 U/ml, failed to augment the proliferative responses to concanavalin A by unfractionated splenocytes or splenic T cells from mice infected for 1 week. In addition, rIL-2 did not modulate the plaque-forming cell response to sheep erythrocytes by splenocytes from these same mice. However, at week 3, rIL-2 in concentrations ranging from 10 to 1,000 U/ml considerably augmented the proliferative response to concanavalin A and plaque-forming cell response to sheep erythrocytes by splenocytes from infected mice. Kinetics studies demonstrated that rIL-2 exerted maximal immunoregulatory activity when added on day 0 or 1 to cultures of splenocytes. In vivo administration of rIL-2, 200 to 20,000 U/day, for 10 days to normal and 3-week-infected mice did not alter the proliferative activity of splenocytes to concanavalin A; 200,000 U of rIL-2 per day actually depressed the proliferative responses of splenocytes from normal and infected mice. In vivo, rIL-2 did not modify delayed-type hypersensitivity responses to sheep erythrocytes or to histoplasmin by normal and infected mice. Moreover, treatment with rIL-2 in vivo did not reduce the number of Histoplasma CFU in spleens of mice. Thus, despite the immunoenhancing effect of rIL-2 in vitro, this lymphokine failed to exert similar effects in vivo.

5-Amino-3-methyl-4-isoxazolecarboxylic acid hydrazide is a non-cytotoxic synthetic isoxazole derivative with considerable immunomodulatory properties demonstrated in in vitro experiments. The aim of this study was to investigate the influence of this compound, depending on the dosage and schedule of treatment, on lymphocyte subsets in non-immunized mice and humoral immune response in SRBC (sheep red blood cells)-immunized mice. An analysis of lymphocyte subsets was carried out by flow cytometry, using specific monoclonal antibodies stained with fluorescein isothiocyanate (FITC) or phycoerythrin (PE). In the SRBC-immunized mice, the influence of the compound on the humoral response was determined, depending on the time of administration relative to the antigen. The number of plaque forming cells (PFC) was determined by a local hemolysis technique in an agar gel. Total and 2-mercaptoethanol resistant serum agglutination titers were defined by active hemagglutination test carried out on microplates. The investigated hydrazide was able to modulate the percentage and absolute number of T lymphocyte subsets in the thymus, and T and B lymphocytes in the peripheral lymphatic organs. It also enhanced humoral immune response in SRBC-immunized mice by increasing the number of cells producing hemolytic anti-SRBC antibodies (PFC) and by augmenting the level of total and 2-mercaptoethanol resistant hemagglutinin. The present study showed modulatory effects of 5-amino-3-methyl-4-isoxazolecarboxylic acid hydrazide on lymphocyte subsets and humoral immune response in mice. This compound could be potentially useful for the treatment of autoimmune diseases, infections or as an adjuvant for boosting the efficacy of vaccines.

African swine fever (ASF) is a highly lethal infectious disease affecting pigs. Although several vaccine formulations have been developed to protect from ASF virus (ASFV) infection, none have yet provided complete protection without side effects or the risk of progressing to chronic infection. mRNA vaccines offer unparalleled advantages in terms of safety and ability to induce immune responses. In this study, we designed six mRNA vaccines encoding key antigenic proteins of ASFV- B602L, CD2V, EP153R, P30, P54, and P72 and combined them into an mRNA cocktail for vaccination in mice and pigs. Our findings suggest that the mRNA cocktail is capable of provoking robust multivalent humoral and cellular immune responses while maintaining a favorable safety profile. Thus, it may serve as a potential approach for controlling ASF transmission, contributing to the ongoing efforts to develop effective and safe ASFV vaccines. The administration of the mRNA cocktail induced both humoral and cellular immune responses in mice and pigs, suggesting a potential for future ASFV vaccine development.IMPORTANCEThis study explores an mRNA vaccine encoding six critical African swine fever virus (ASFV) antigens (B602L, CD2V, EP153R, P30, P54, P72), demonstrating its ability to induce robust humoral and cellular immune responses in both mice and pigs. This innovative approach serves as a significant advancement in ASFV vaccine development by addressing safety and efficacy concerns. The findings suggest that the mRNA cocktail developed in this study represents a step forward in ASFV vaccine research and development. This strategy holds promise for contributing to ASFV control by offering possibly safer and more effective alternatives to conventional vaccines. This could significantly impact ASF management and prevention strategies globally, ultimately benefiting animal health and reducing economic losses.

In order to develop an experimental DNA vaccine for the prevention and treatment of hepatitis B virus infection, hepatitis B virus surface antigen (HBsAg) DNA was subcloned into an E. coli-eukaryotic cell shuttle vector and was expressed in the Baculovirus expression system. Intramuscular, intradermal, and intraperitoneal injections of 30 microg of the plasmid DNA expressing HBsAg induced humoral and cellular immune responses in ICR mice. The first IgG antibodies were detected after ten days and specific IgG antibody titers peaked after two months of a single intramuscular DNA injection. Anti-HBs antibody titers gradually increased and peaked at four months following intradermal DNA injection, and in case of intraperitoneal injection they peaked at seven months. Generation of HBs-specific helper T lymphocytes was also investigated through the production of interleukin-2 by T helper cells. Boosting effects of HBs DNA were investigated without much results. In general, DNA-mediated HBs immunization induced humoral and cellular immune responses in mice that appears to simulate immune responses in human during the course of HBV vaccination.

E-55+ murine leukemia virus infection of both progressor (BALB) and long term nonprogressor (C57BL) mouse strains is characterized by an acute and a persistent phase of infection. During the acute phase, progressor strains require CD8+ T cells to decrease virus burden, whereas the long term nonprogressor strains do not. In the present studies the immune response in BALB and C57BL mice during the acute phase of E-55+ murine leukemia virus infection was examined. The results demonstrate that BALB mice produce both IL-4 and IFN-gamma, in contrast to C57BL mice, which produce only IFN-gamma. In BALB mice, IL-4 production results in the absolute requirement for CD8+ T cells to reduce the virus burden during the acute phase of infection. The anti-virus immune response in these mice is IFN-gamma dependent. On the other hand, C57BL mice do not produce IL-4 and, in the absence of both CD8+ T cells and IFN-gamma, still generate an effective anti-virus immune response. Genetic studies suggest that these distinct immune responses are regulated by more than one non-MHC-linked gene. Two candidate regions that may encode this gene(s), located on chromosomes 7 and 19, respectively, were identified by recombinant inbred strain linkage analysis.

miR‐107‐5p ameliorates neurological damage, oxidative stress, and immune responses in mice with Alzheimer's disease by suppressing the Toll‐like receptor 4 (TLR4)/nuclear factor‐kappaB(NF‐κB) pathway

Objective To investigate the development strategy of novel T cell based vaccine against HCV infection. Methods BALB/c mice were primed with pSCK-based DNA vaccine and boosted with type 5 adenoviral vector-based vaccine, which expressed the structural proteins (Core, E1 and E2) derived from a Chinese HCV patient (genotype 1b, Hebei strain). Enzyme linked immunospot assay (ELISPOT) and intracellular cytokine staining (ICS) were used to analyze the elicited antigen-specific immune responses and the efficacy of cross-protection. Results Immunization of mice with the prime-boost vaccination strategy elicited stronger T cell immune responses against multiple HCV antigens than using the DNA vaccines alone, especially the IFN-γ-secreting T cell responses against E1 protein as indicated by ELISPOT assay. ICS data indicated that the prime-boost regimen elicited more TNF-α-producing CD4+ and IFN-γ-producing CD8+ T cells against E1 protein and high levels of IFN-γ-producing CD4+ and CD8+ T cells against E2 protein in comparison with immunization with DNA vaccines. Moreover, the prime-boost vaccination was capable of eliciting effective cross-protection in a surrogate challenge model based on a recombinant heterologous HCV (JFH1, 2a) vaccinia virus. Conclusion The prime-boost vaccination using DNA and rAd5-based vaccine expressing HCV structural antigens induced significant cellular immune response and cross-protection in mice, suggesting the possibility of using it as a promising T cell based vaccine against HCV infection. Key words: Hepatitis C virus; DNA vaccine; rAd5-based vaccine; T cell response; Prime-boost vaccination

In previous studies, we demonstrated that CRA and FRA recombinant proteins, used for diagnosis of Chagas' disease, elicited a humoral immune response in susceptible and resistant mice. To understand better the immune response to these proteins, we have evaluated, the cellular immune response in CRA- and in FRA-immunized BALB/c and C57BL/6 mice. A specific cellular lymphoproliferative response was observed in both strains of mice. Spleen cell cultures mainly from CRA-immunized C57BL/6 and FRA-immunized BALB/c mice produced high levels of IFN-y, indicating the induction of a Type 1 immune response. Regarding the T cell subsets, CD4+ T cells were the major source of IFN-y in CRA- and FRA-immunized mice. These results suggest that CRA and FRA are important immunogens in inducing a Type 1 immune response and that they may be considered as potential vaccine antigens.

ADP-ribosylation factor 1 (HcARF1) is one of the Haemonchus contortus (H. contortus) excretory/secretory proteins involved in modulating the immune response of goat peripheral blood mononuclear cells (PBMC). Here, we evaluated the immunogenic potential of recombinant HcARF1 (rHcARF1) against H. contortus infection in Institute of Cancer Research (ICR) mice. Briefly, rHcARF1 was entrapped in poly (D, L-lactide-co-glycolide) (PLGA) and chitosan (CS) nanoparticles (NP) and injected into mice as a vaccine. Fifty-six ICR mice were assigned randomly into seven groups, with eight animals in each group, and they were vaccinated subcutaneously. At the end of the experiment (14th day), the blood and the spleen were collected from euthanized mice to detect lymphocyte proliferation, cytokine analysis, and the production of antigen-specific antibodies. Scanning electron microscope was used to determine the size, morphology, and zeta potential of nanoparticles. Flow cytometry was performed, which presented the increase percentages of CD4+ T cells (CD3e+CD4+), CD8+ T cells (CD3e+CD8+) and dendritic cells (CD11c+CD83+, CD11c+CD86+) in mice vaccinated with rHcARF1+PLGA NP. Immunoassay analysis show raised humoral (Immunoglobulin (Ig)G1, IgG2a, IgM) and cell-mediated immune response (Interleukin (IL)-4, IL-12, and IL-17, and Interferon (IFN)-γ) induced by rHcARF1+PLGA NP. Experimental groups that were treated with the antigen-loaded NP yield higher lymphocyte proliferation than the control groups. Based on these results, we could propose that the rHcARF1 encapsulated in NP could stimulate a strong immune response in mice rather than administering alone against the infection of H. contortus.

The effect of different psychological profiles and timings of stress exposure on humoral immune response