The Immunomodulatory Effects of Phellodendri Cortex Polysaccharides on Cyclophosphamide-Induced Immunosuppression in Mice

Cyclophosphamide is a commonly used anticancer drug, and immunosuppression is one of the most common side effects. How to recover the immunological function is important for cyclophosphamide-treated patients. In the present study, Phellodendri Cortex polysaccharides (CPP) could enhance the proliferation of mouse spleen lymphocytes in vitro. The immunoregulatory function of CPP was then investigated in cyclophosphamide-induced immunosuppressed mice. In CPP-treated groups, mice were orally treated with CPP at doses of 1, 0.5, and 0.25 g/kg bodyweight from 1 to 11 d, respectively. The cyclophosphamide was administrated in CPP and cyclophosphamide groups from 12 to 14 d. In the cyclophosphamide and normal control groups, the mice received equal volume of saline from 1 to 14 d. The results showed that CPP (1 g/kg) could significantly increase the bodyweight of mice, even during cyclophosphamide treatment. The organ coefficients of the spleen and thymus were recovered by CPP treatment. CPP upregulated the contents of cytokines (IL-2, IL-6, IFN-γ, and TNF-α) in serum, which were downregulated by cyclophosphamide. The mRNA levels of these cytokines were also elevated by CPP treatment in the spleen. Cyclophosphamide upregulated the expressions of NF-κB p65, TLR4, and MyD88, suggesting that the NF-κB signaling pathway was activated by cyclophosphamide. After CPP treatment, it was recovered to normal level. These results indicated that CPP alleviated the cyclophosphamide-induced immunosuppression.

Paulownia tomentosa flower polysaccharide as an effective immunopotentiator to enhance immune responses for Newcastle disease vaccine in mice

To investigate the immunomodulatory activity and explore the mechanism of Paulownia tomentosa flower polysaccharides (PTFP). PTFP was orally administrated to mice for seven successive days before and after Newcastle disease vaccination. The results demonstrated that compared with the vaccine control (VC) group, PTFP enhanced the inhibition of hemagglutination assay antibody titers, promoted the antigen-specific immunoglobulin (Ig)G, IgG1, IgG2a, and IgG2b antibodies responses, enhanced proliferation of spleen T and B lymphocytes, increased the secretions of interferon-γ and interleukin-10 cytokines of spleen lymphocytes, and promoted the activation of natural killer cells. Therefore, PTFP, as an effective immunopotentiator, could induce a mixed T-helper (Th)1 and Th2 immune responses and an innate immune response.

Tim-3ScFv-Transforming Lactobacillus Inhibits Transplanted Tumour of Renal Cell Carcinoma

Background T cell immunoglobulin-3(Tim-3) is an immune checkpoint molecule; Tim-3 antibody is suitable for treating malignant renal tumours. However, Tim-3 antibody drugs are expensive, which limits their application. To overcome the disadvantages of expensive immunotherapeutic drugs, Lactococcus lactis was used as the host bacteria to express Tim-3 single-chain antibody in the intestine, and its promotion of the mouse immune system and inhibitory effects on the transplanted tumour of kidney cancer in mice were tested. Methods Molecular cloning technology was used to construct plasmids pLAN-CTB-Tim3scFv and pLAN-Tim3scFv, which were transformed into Lactococcus lactis. The expression of the transformed bacteria was analysed using western blotting, and the immune activity of secreted proteins of the transformed bacteria was detected using ELISA in vitro. A subcutaneous transplanted tumour model of renal adenocarcinoma was constructed in RAG mice, and the promoting effect of transforming bacteria on the activation of mouse spleen lymphocytes, and its inhibitory effect on transplanted tumours in mice was analysed. Results (1) Transformed Lactococcus lactis NZ -CTB-Tim3scFv and NZ -Tim3scFv, which secrete CTB-Tim3scFv and Tim3scFv single-chain antibodies, were successfully constructed. (2) CTB-Tim3scFv secreted by NZ-CTB-Tim3scFv transformed bacteria showed immunological activity. (3) Compared with the NZ-Tim3scFv and NZ-Vector groups, the characteristics of the subgroups of splenic lymphocytes in the NZ-CTB-Tim3scFv group had a higher proportion of CD3+CD4+, CD3+CD8a+, and CD3+CD69+ cells. Ki67 and CD31 expression in the NZ-CTB-Tim3scFv group was significantly reduced. The tumour volume of the NZ-CTB-Tim3scFv group increased the least, and was statistically different from that of the other two groups. Conclusions The Tim-3 single-chain antibody gene was successfully constructed and transformed in Lactococcus lactis. After feeding mice NZ-CTB-Tim3scFv transforming Lactobacillus, the CTB-Tim3scFv secreted by the transforming Lactobacillus promoted the proliferation and activation of spleen lymphocytes and inhibited volume growth, cell proliferation, and angiogenesis of the tumour in mice. In summary, apply transgenic lactobacillus secreting CTB-Tim-3scFv, to perform its role in anti-tumor immunotherapy through oral approach, is low cost and convenient. It is expected to become a new way of immunotherapy for renal cell carcinoma.

Xenogeneic lymphocytic RNA stimulates skeletal muscle regeneration

Objective: to find evidence of the existence of distant lymphocytic RNA control of physiological myogenesis as a way to control the muscle tissue regeneration process.Materials and methods. The study was conducted on male Wistar rats, n=33. In the first part of the experiment, 12 rats were subjected to regular 40-day physical activity (swimming), half of them were intraperitoneally injected 4 times with total RNA isolated from pig spleen lymphocytes at 30 days of age; 6 rats made up the intact control group. In histological preparations of different skeletal muscle groups, the width and cross-sectional area of muscle fibers, the area of nuclei, and the number of myocytes and myosatellite cells were evaluated. In the second part of the experiment, 15 intact rats were injected with the studied xenogeneic RNA and the amounts of ribonucleic acids in peripheral blood lymphocytes, spleen lymphocytes, and skeletal muscles were determined 2 hours and 24 hours after injection.Results. After the 40- day physical activity, the width of the fibers and the area of myocyte nuclei in the skeletal muscles increased; the absolute number of myosatellite cells and the area of their nuclei did not change. After administration of xenogenic RNA in the trained rats, in addition to an increase in the thickness and cross-sectional area of muscle fibers, the absolute number of myosatellite cells in m. biceps femoris, in m. triceps brachii, and in m. pectoralis major increased 1.4-fold, 1.3-fold, and 1.4-fold, respectively; the area of myosatellite nuclei increased on average by 7%. In intact rats, two hours after xenogeneic RNA injection, the amount of RNA in skeletal muscles remained unchanged, it increased by 19% in spleen lymphocytes, and by 16% in peripheral blood lymphocytes. At 24 hours, the RNA amount in the lymphocytes remained significantly higher than the control values, while in the muscle tissue, it didn’t differ from the control.Conclusion. Xenogeneic lymphocytic RNA stimulates physiological myogenesis by activating myosatellite cell proliferation.

Immunomodulatory Effect of Pinus Armandi Franch Seed Oil

Pinus armandi franch is a special local plant in China and its seed oil is a nice resource of linoleic acid (LA) and linolenic acid. This study investigated the effects of enriching the mouse diet with the ω-6 and ω-3 polyunsaturated fatty acids (PUFA), LA (18:2 ω-6) and α-linolenic acid (18:3 ω-3) different ratios (ω-6/ω-3) on total lipids in Pinus armandi franch seed oil (PAFO), and immune evaluation indexes in plasma, lymph nodes, and spleen from isolated immune cells. Kunming mice were fed a commercial chow. PAFO with a certain ratio (ω-6/ω-3) could promote the proliferation of spleen lymphocytes induced by ConA in mice and the optimal ratio was 4 or 6. PAFO with different ratios (ω-6/ω-3) could increase the spleen index and thymus index and there were significant differences between PAFO group and control group (p < 0.01). The results indicate that the ratio of ω-6 PUFA: ω-3 PUFA is 4 or 6 improve some of the indices of the mice immune status and that a moderate increase in the level of ω-6 PUFA in the mice diet does not suppress the immune system.

Development of lymphocyte subpopulations in local breed chickens

Background and Aim: Local breeds of chicken are known to have relatively higher disease resistance to many endemic diseases and diseases that are highly virulent in commercial chickens. This study aimed to address the lymphocyte subpopulations in three constitutive immune system organs (thymus, bursa of Fabricius, and spleen) in 30, 8-week-old, male local breed chickens. Materials and Methods: The T (CD3+) and B lymphocytes (Bu-1+) were identified through one-color, direct immunofluorescent staining of the thymus, bursa, and spleen lymphocytes. Likewise, two-color, direct immunofluorescent staining was performed to identify the CD4- and/or CD8-defined T lymphocytes. The proportions of T and B lymphocytes and CD4- and/or CD8 defined chicken lymphocyte subsets in lymphoid suspensions prepared from the thymus, bursa, and spleen were determined by flow cytometry. Results: CD3+ cells, particularly those positive for CD4+CD8–, were dominant in the thymus, whereas cells expressing the Bu-1 marker were predominant in the bursa of Fabricius. The proportion of T and B cells was almost equal in the spleen, with more cells expressing the CD4–CD8+ marker in the red pulp. Conclusion: These findings indicate that local breeds of chicken could serve as a reliable model for studying the immune system of commercial light chicken breeds, due to the similarity in the presence and the distribution of the immune cells.

Microwave-Assisted Enzymatic Extraction of Flavonoids from Armeniaca mume Sieb. Blossom and Their Immunomodulating Effect in Mice with DSS-Induced Colitis

Armeniaca mume Sieb. blossom is among the traditional Chinese edible flowers, and it is widely used in the food and pharmaceutical industries. Flavonoids are among the most abundant bioactive compounds in A. mume Sieb. blossom. However, the research on the extraction of flavonoids from A. mume Sieb. blossom and their immunomodulating function is insufficient. In this study, we developed a microwave-assisted enzymatic extraction of flavonoids from A. mume Sieb. blossom (FAMB) and explored their immunomodulating effect on mice with dextran sulfate sodium salt-induced colitis. The results showed that the optimum parameters for microwave-assisted enzymatic extraction of FAMB were as follows: cellulase: 2.0%; microwave power: 200 W; microwave action time: 5 min; and enzymatic hydrolysis time: 50 min. FAMB significantly promoted the lymphocyte proliferation and natural killer (NK) cell killing activity in colitis mice, and increased the concentrations of TNF-α, IFN-γ, and IL-2 in serum. FAMB also significantly reduced the apoptosis of spleen lymphocytes in these mice. These results demonstrated that the microwave-assisted enzymatic method could significantly improve the yield and efficacy extraction of FAMB. FAMB showed a good immunomodulation effect on colitis mice.

Evaluation of Immune Function of Mycelium Fermentation of Hirsutella sinensisin In Vitro and In Vivo

In order to confirm whether HSFL (the fermented liquor of mycelia of Hirsutella sinensis) still contains cordycepin, Cordyceps polysaccharide and other functional components, and has the functions of anti-oxidation, tumor inhibition and immunity enhancement, the biological activity of HSFL In Vivo and in vitro was studied in this study. The transformation ability of mouse spleen lymphocytes induced by ConA, the activity of NK cells in mouse spleen, the delayed allergic reaction induced by DNFB and the phagocytosis of chicken red cells by mouse peritoneal macrophages were studied. The results showed that when the concentration of HSFL was 0.5972 mg/kg and 1.1944 mg/kg, the transformation of lymphocytes induced by ConA and the activity of NK cells were significantly increased. HSFL also can significantly improve DNFB induced anaphylaxis in mice and phagocytosis of chicken erythrocytes by peritoneal macrophages in mice when the dose of HSFL is 50 mg/kg and 75 mg/kg, indicating that HSFL has the biological activity of enhancing immunity in vitro and In Vivo.

RESEARCH OF RESTORATION PROCESSES AFTER INDUCTION OF LYMPHOPENIA BY CYCLOPHOSPHANE IN AN EXPERIMENTAL MODEL ON MOUSE

Objective: to evaluate the dynamics of T-cell recovery after exposure to cyclophosphamide (CP). CP was injected at a dose of 125 mg/kg, followed by killing of mice on days 0, 10, 20, 30, 60 and determination of the subpopulation composition by flow cytometry. It was shown that thymocytes are more vulnerable to the action of the CP than spleen lymphocytes. Cell restoration in the thymus begins earlier with delayed recovery of spleen lymphocytes. Due to CP, there is an increase in the conversion of the naive T-cells into central memory T-cells with a relative accumulation of the latter.