玉米赤霉烯酮饲粮添加蒙脱石对断奶仔猪脾脏和外周血免疫指标的影响

选择(28±1)日龄、平均体重为(8.84 ±0.21)kg的健康三元(斯格×长×大)杂交断奶仔猪(公母各半)18头,研究1 mg/kg玉米赤霉烯酮(zearalenone,ZEA)污染饲粮对断奶仔猪脾脏和外周血淋巴细胞增殖率和IL-2产量的影响,同时评价改性蒙脱石的脱毒效应.将试验仔猪按照体重随机分为3个处理(公母各半),仔猪采用试验笼单独饲养.试验处理为:Contr.=基础饲粮;ZEA+CZ0=基础饲粮+1 mg/kg ZEA;ZEA+ CZ4=基础饲粮+1 mg/kg ZEA+4g/kg改性蒙脱石.预饲期7d,正式期22 d.结果表明:1)与对照组相比,饲粮添加1 mg/kg ZEA显著降低断奶仔猪脾脏相对重量、外周血和脾脏淋巴细胞增殖率以及IL-2产量(P＜0.05).2)1 mg/kg的ZEA处理饲粮添加4 g/kg改性蒙脱石能够显著改善ZEA诱导的脾脏和外周血淋巴细胞增值率以及IL-2的改变(P＜0.05).结果揭示,1 mg/kg的ZEA足以影响断奶仔猪脾脏的生长发育及其细胞免疫功能,添加4 g/kg改性蒙脱石具有显著的脱毒效应,以上结果对指导动物生产和人类健康具有重要的借鉴意义.     

Bovine-associated staphylococci and mammaliicocci trigger T-lymphocyte proliferative response and cytokine production differently

We examined whether distinct staphylococcal and mammaliicoccal species and strains trigger B- and T-lymphocyte proliferation and interleukin (IL)-17A and interferon (IFN)-γ production by peripheral blood mononuclear cells in nulliparous, primiparous, and multiparous dairy cows. Flow cytometry was used to measure lymphocyte proliferation with the Ki67 antibody, and specific monoclonal antibodies were used to identify CD3, CD4, and CD8 T lymphocyte and CD21 B lymphocyte populations. The supernatant of the peripheral blood mononuclear cell culture was used to measure IL-17A and IFN-γ production. Two distinct, inactivated strains of bovine-associated Staphylococcus aureus [one causing a persistent intramammary infection (IMI) and the other from the nose], 2 inactivated Staphylococcus chromogenes strains [one causing an IMI and the other from a teat apex), as well as an inactivated Mammaliicoccus fleurettii strain originating from sawdust from a dairy farm, and the mitogens concanavalin A and phytohemagglutinin M-form (both specifically to measure lymphocyte proliferation) were studied. In contrast to the “commensal” Staph. aureus strain originating from the nose, the Staph. aureus strain causing a persistent IMI triggered proliferation of CD4⁺ and CD8⁺ subpopulations of T lymphocytes. The M. fleurettii strain and the 2 Staph. chromogenes strains had no effect on T- or B-cell proliferation. Furthermore, both Staph. aureus and Staph. chromogenes strains causing persistent IMI significantly increased IL-17A and IFN-γ production by peripheral blood mononuclear cells. Overall, multiparous cows tended to have a higher B-lymphocyte and a lower T-lymphocyte proliferative response than primiparous and nulliparous cows. Peripheral blood mononuclear cells of multiparous cows also produced significantly more IL-17A and IFN-γ. In contrast to concanavalin A, phytohemagglutinin M-form selectively stimulated T-cell proliferation.     

Effects of low dose oxymatrine on mouse lymphocyte proliferation stimulated by Con A

Objective To investigate the effects of low dose Oxymatrine (OMT) on mouse lymphocyte proliferation stimulated by Con A making use of fluorescence dyestuff (CFDA-SE). Methods CFDA-SE staining and flow cytometry were used to detect the fluorescence intensity of lymphocytes after stimulated by polyclonal stimulators Con A and OMT. Then, related software was used to analyze the effects of OMT on mouse lymphocyte proliferation. Results After cultured for 48 h, CFSE fluorescence could be detected by cytometer, filial generation peaks did not appear in control group, which indicated that lymphocytes did not proliferate. Three peaks were obviously detected in Con A group which indicated that Lymphocytes divided after 48 h stimulated by Con A compared with the halving of the fluorescence intensity of control group. In groups with Con A and OMT treated. Primary generation peaks are all lower while filial generation peaks are significantly higher than groups with Con A treated only. This indicated OMT obviously promote lymphocyte proliferation. After cultured for 72 h, the fluorescence intensity changes between all groups are consistent with those of cultured for 48 h. Analyzed with CELLQuest software, it is shown that OMT could promote lymphocyte proliferation in 16, 8, 4 and 2 μg/mL respectively. Conclusions 1)CFDA-SE dyeing and flow cytometer were both reliable tools to analyse lymphocyte proliferation; 2) lower dosage of OMT could promote the proliferation of lymphocyte as a immunopotentiator. Correspondence     

Comparative analysis of one-particle mathematical models in materials science and oncology. II. Qualitative study of the equations

A system of two nonlinear first-order differential equations is analyzed that describe the dynamics of the interaction between lymphocytes and tumor cells in the body. The model utilized takes into account the effect of a magnetic field. A concept is formulated concerning the analogy between the behavior of microparticles (atoms, molecules) in a medium with traps (in materials science) and the behavior of lymphocytes in living tissue containing tumor cells (in oncology). It is established that the tools of the qualitative theory of differential equations (analogous to those applicable to oncology) can be effective in analysis of transport and transformations in materials science. __________ Translated from Poroshkovaya Metallurgiya, Nos. 1–2(447), pp. 92–98, January–February, 2006.     

ABV方案化疗对艾滋病相关晚期卡波氏肉瘤外周血CD4淋巴细胞的影响

Objective: The combination of highly active antiretroviral therapy (HAART) and chemotherapy with ABV regimen (doxorubicin, bleomycln and vincristine) is a promising approach for the treatment of advanced HIV-related Kaposi's sarcoma (KS). Here we analyzed the relationship between the CD4 lymphocyte cell count and the clinical response to chemotherapy.Methods: The 176 HIV infected patients with advanced KS who failed to respond to prior HAART were selected. All these patients were then preceded to chemotherapy with ABV regimen which was administered at 3 weekly intervals for 6 cycles.For each patient CD4 cell count was done before starting chemotherapy and after finishing 6 cycles of chemotherapy. The difference of CD4 cell counts pre chemotherapy and post chemotherapy was compared with the clinical progress of the patients after 6 cycles of chemotherapy. Results: The overall clinical remission was shown in 93.7% patients. Progressive disease (PD) and no change in clinical condition (NC) was shown in 6.3% patients. The increase in CD4 cell count post chemotherapy was found in 89.8% patients and the decrease in CD4 cell count was seen in 10.2% patients. The difference of the mean CD4call counts for patients in group CR + PR (complete relief + partial relief) before and after chemotherapy was highly significant.The difference of the mean CD4 cell counts for patients in group NC + PD before and after chemotherapy was not significant.The difference in CD4 cell counts in CR + PR and NC + PD groups before and after chemotherapy was highly significant.Conclusion: The HIV related KS patients on HAART benefit from the chemotherapy as it increases the CD4 cell count and it has positive impact on clinical remission of KS.     

不同来源Lfcin对小鼠脾淋巴细胞增殖及分泌细胞因子影响的对比研究

本文以牛乳铁蛋白素Lfcin B和人乳铁蛋白素Lfcin H为研究对象,研究其对小鼠脾淋巴细胞增殖及分泌细胞因子的影响并比较两者的差异及相关性。采用CCK-8法检测不同浓度的Lfcin单独作用对小鼠脾淋巴细胞增殖的影响及不同质量浓度Lfcin与丝裂原共同作用对小鼠T、B淋巴细胞增殖作用的影响;ELISA法测定IL-2、IL-4和TNF-α的含量。结果显示:Lfcin B和Lfcin H能显著促进脾淋巴细胞增殖及增强Con A诱导的T淋巴细胞的增殖和LPS诱导的B淋巴细胞的增殖,并且能显著促进脾淋巴细胞分泌IL-2、IL-4、TNF-α(P0.01)。研究结果表明:Lfcin能有效的促进脾淋巴细胞的增殖,并与Con A、LPS对T、B淋巴细胞的增殖有协同刺激作用,其机制可能是通过促进细胞因子的分泌发挥免疫增强作用。     

Lymphocyte functions in overconditioned cows around parturition

The objective of the study was to evaluate the relationships between body condition and lymphocyte functions in periparturient dairy cows. Thirty days before expected calving, 21 Holstein cows were categorized as thin (n = 6), medium (n = 8), or overconditioned (n = 7) based on body condition score (BCS). Blood samples were collected on 21, 14, 7, and 3 d before calving and on d 3, 7, 14, 21, 28, and 35 after parturition. An aliquot of blood was used to determine plasma nonesterified fatty acids (NEFA) and glucose. At 14 and 7 d before, and 14 and 35 d after calving, a second aliquot of blood was used to assess peripheral blood mononuclear cell (PBMC) functions: DNA synthesis, immunoglobulin (Ig) M, and interferon-gamma (IFN-γ) secretion after mitogen stimulation. During the experiment, all 21 cows showed a decline in BCS. Overconditioned cows lost significantly more BCS than thin cows. After calving, overconditioned cows had higher plasma NEFA compared with thin and medium cows. Conversely, plasma glucose never differed between the 3 categories of cows. Regardless of BCS, DNA synthesis and IgM secretions were significantly lower in PBMC isolated on 7 d before calving compared with those recorded 14 and 35 d after parturition. Conversely, PBMC from the 21 cows did not show any change of IFN-γ secretion during the experimental period. Taking into consideration the BCS categories, PBMC isolated from overconditioned cows presented lower IgM secretion compared with thin cows on d 14 and 35 after calving. Furthermore, PBMC isolated from overconditioned cows secreted less IFN-γ compared with thin and medium cows on d 7 before calving. The DNA synthesis of PBMC stimulated with the 3 mitogens did not differ between the 3 categories of cows. In conclusion, immunodepression occurring in cows around calving would be particularly evident in overconditioned cows.     

Effect of different fatty acids on the proliferation and cytokine production of dairy cow peripheral blood mononuclear cells

During the transition period, dairy cows often experience negative energy balance, which induces metabolic and immunological disturbances. Our previous work has shown a relationship between the inhibition of immune functions and increased blood nonesterified fatty acid (NEFA) levels. In this study, we evaluated the effect of 11 fatty acids (palmitoleic, myristic, palmitic, stearic, oleic, linoleic, docosahexaenoic, conjugated linoleic, lauric, eicosapentaenoic, and linolenic acids) as well as a mix that represented the NEFA profile observed during the transition period at different concentrations (0, 50, 100, and 250 µM) on proliferation and cytokines secretion of lymphocytes. To assess lymphoproliferation, peripheral blood mononuclear cell (PBMC) from 5 healthy cows (166–189 d in milk) were isolated, stimulated with the mitogenic lectin concanavalin A (ConA), incubated for 72 h with or without fatty acids, and subjected to flow cytometry analysis. Our results showed that all fatty acids, except lauric acid, significantly reduced proliferation of PBMC in a dose-dependent manner. The most detrimental effect was observed with conjugated linoleic and stearic acids, where proliferation of PBMC was already inhibited at the lowest dose (50 µM). For cytokine secretion, we found that levels of IL-4 in culture supernatant of ConA-stimulated PBMC were reduced after a 24-h exposure to the lowest dose (50 µM) of oleic and palmitoleic acids. A dose of 100 µM of eicosapentaenoic acid, NEFA mixture, and myristic acid was necessary to observe a reduction in IL-4 levels. The PBMC also showed a decrease in the secretion of IFN-γ in response to lauric, linolenic, palmitoleic, and stearic acids at 50 µM and myristic acid at 100 µM. Overall, polyunsaturated fatty acids were more potent inhibitors of cytokine secretions than saturated fatty acids. In addition, we detected an inverse relationship between the melting points of fatty acids and their ability to inhibit IL-4 and IFN-γ secretions, as evidenced by greater inhibition with low–melting point fatty acids. Overall, our study confirmed that NEFA have a negative effect on some lymphocyte functions, and that their inhibitory effect on cytokine secretions increases with the degree of unsaturation.     

WT1蛋白CTL表位肽基因载体的构建与鉴定

目的构建WT1(Wilms’tumor gene 1)蛋白CTL表位肽基因载体,并检测其在293T细胞中的转录。方法设计分别含WT1-126肽、WT1-235肽以及这两种肽的基因载体,并加入Th通用表位Pan-DR-Th(PADRE),应用蛋白酶体切割软件PAProC和NetChop优化各表位和间隔序列,DNA疫苗在线预测工具DyNAVacS优化真核密码子后,人工合成核苷酸序列,分别插入pUC57载体,构建pUC57-WT1质粒,测序鉴定后,酶切回收各目的片段,亚克隆至真核表达载体pcDNA3.1(+)中,构建重组真核表达质粒,转染293T细胞,RT-PCR检测各目的基因在293T细胞中的转录。采用无内毒素质粒大量提取试剂盒提取各重组质粒,采用紫外分光光度计测定质粒的纯度和浓度。结果各重组真核表达质粒经双酶切及测序鉴定证实构建正确;重组质粒携带的目的基因可在293T细胞中成功转录;各重组质粒DNA的纯度均合格,浓度在864.6～883.9μg/ml之间。结论成功构建了WT1蛋白CTL表位肽基因载体,并能在真核细胞中正常转录,为下一步在小鼠体内探讨特异性不同的CTLs群发挥抗肿瘤作用的机制奠定了基础。     

Characterization of genetic polymorphism of the bovine lymphocyte antigen DRB3.2 locus in Kankrej cattle (Bos indicus)

Bovine lymphocyte antigen DRB 3.2 (BoLA-DRB3.2) gene encodes for the beta chain of the major histocompatibility complex (MHC) class II molecule in cattle, which is a glycoprotein present on the surface of antigen-presenting cells. This locus shows extensive polymorphism in it. The objective of the present study was to genotype the BoLA-DRB3.2 locus in Kankrej cattle (n = 50) by PCR-RFLP. Bovine DNA was isolated from aliquots of whole blood. Primers specific for exon 2 of the bovine lymphocyte antigen (BoLA)-DRB3 gene were used to amplify the region. The 304-bp amplified product of the DRB3 gene was separately digested with restriction endonucleases RsaI, BstYI, and Hae III. Twenty-four BoLA-DRB 3.2 alleles were identified with frequencies ranging from 1 to 22.0%. Twenty-one alleles of the total 24 alleles were similar to those reported earlier; 3 alleles were new and had not been reported previously. The allele BoLA-DRB3.2*34 occurred at the highest frequency of 22% (approx.) in the Kankrej animals studied. Six alleles (BoLA-DRB3.2 *34, *15, *06, *20, *37, and *20) accounted for almost 71% of the total alleles observed to be present in the Kankrej animals. All the new alleles observed were present at frequencies of 1%. The results obtained in the present study demonstrated that the BoLA DRB3.2 locus is highly polymorphic in the Kankrej cattle.