Adiloğlu Ali Kudret, Gönülateş Nurettin, Işler Mehmet, Senol Altuğ
Department of Medical Microbiology, Ankara Education and Research Hospital, Ankara, Turkey.
Mikrobiyol Bul. 2013 Apr;47(2):273-81. doi: 10.5578/mb.4709.
The systemic effects of bioactive peptides which are produced by the fermentation of milk via the microorganisms found in kefir have been the subject of interest in recent years. Bioactive peptides activate innate immunity by stimulating macrophages, increasing phagocytosis, augmenting NO and cytokine production and boosting the lumen levels of IgG and IgA+ B-lymphocytes. The aim of the present study was to determine the serum cytokine profiles of healthy volunteers after kefir consumption to evaluate helper T (TH) cell polarization and to bring out the effects on native and allergic immune responses. The study was designed as a prospective and self-controlled study. A total of 18 healthy volunteers (age range: 20-40 yrs, mean age: 35.5 ± 7.38 yrs) from a university hospital staff were recruited to the study, with the approval of ethical board and informed consent. The body mass indices of all participants were between normal range (20.10-25.70 kg/m2). After two weeks of a diet free from fermented products, the participants consumed 200 mL kefir daily, for six weeks. Kefir product was prepared by using kefir starter culture (Danisco Biolacta Sp - 05223B 10001, Poland) which contains Lactobacillus spp., Leuconostoc spp., Lactococcus lactis ssp. lactis and Streptococcus termophilus, an overnight incubation at 26°C, and consumed freshly. Fasting blood samples of subjects were collected just before kefir use (0th week), at the end of the 3rd and 6th weeks of kefir consumption, and three weeks after cessation of kefir usage (9th week). Serum TNF-a, IL-1, IL-5, IL-8 and TGF-β levels were measured by using commercial ELISA kits (BioSource, Belgium and Invitrogen, USA). Hemoglobin, serum creatinine and ALT levels of all subjects were also determined for follow-up. All volunteers completed the study period without any problem and declared no complaint. Hemoglobin, creatinine and ALT levels did not change with kefir consumption. Serum IL-8 levels were decreased at 3rd and 6th weeks (p< 0.001) and were at low levels at 9th week (p= 0.005) when compared with baseline levels (0th week). Serum IL-5 levels were increased at 3rd week (0th-3rd weeks; p= 0.01) and decreased by a rebound effect at 9th week (6th-9th week p= 0.003). TNF-α levels were increased with kefir consumption (p= 0.046) but the increase was insignificant in paired comparisons and the level was borderline between 0th and 6th weeks (p= 0.013). IL-5 and TNF-α levels returned to their original levels (0th week) at 9th week. Levels of the other cytokines (IL-1 and TGF-β) did not change significantly with kefir usage. These results indicated that kefir use increased polarization of the immune response towards TH1 type and decreased TH2 type response and accordingly allergic response. The decrease in IL-8 level due to kefir use, might control the inflammatory response by suppressing neutrophil chemotaxis and activation. On the other hand it was also concluded that increased IL-5 might stimulate secretory IgA at gastrointestinal mucosa leading to a more efficient immune response in the intestinal lumen.
近年来,开菲尔中发现的微生物发酵牛奶产生的生物活性肽的全身效应一直是人们感兴趣的课题。生物活性肽通过刺激巨噬细胞、增加吞噬作用、增强一氧化氮和细胞因子的产生以及提高管腔中IgG和IgA+B淋巴细胞水平来激活先天免疫。本研究的目的是确定健康志愿者饮用开菲尔后的血清细胞因子谱,以评估辅助性T(TH)细胞极化,并揭示其对天然免疫和过敏免疫反应的影响。该研究设计为前瞻性自我对照研究。在伦理委员会批准并获得知情同意后,从一家大学医院的工作人员中招募了18名健康志愿者(年龄范围:20 - 40岁,平均年龄:35.5±7.38岁)。所有参与者的体重指数均在正常范围内(20.10 - 25.70kg/m²)。在两周无发酵产品饮食后,参与者每天饮用200mL开菲尔,持续六周。开菲尔产品是使用含有乳酸杆菌属、明串珠菌属、乳酸乳球菌乳酸亚种和嗜热链球菌的开菲尔发酵剂培养物(丹麦丹尼斯克生物乳酸公司 - 05223B 10001,波兰)制备的,在26°C下过夜培养,并新鲜饮用。在饮用开菲尔前(第0周)、饮用开菲尔第3周和第6周结束时以及停止饮用开菲尔三周后(第9周)采集受试者的空腹血样。使用商业ELISA试剂盒(比利时BioSource公司和美国Invitrogen公司)测量血清TNF - α、IL - 1、IL - 5、IL - 8和TGF - β水平。还对所有受试者的血红蛋白、血清肌酐和ALT水平进行了随访测定。所有志愿者均顺利完成研究期,且无任何不适主诉。血红蛋白、肌酐和ALT水平在饮用开菲尔后未发生变化。与基线水平(第0周)相比,血清IL - 8水平在第3周和第6周时降低(p < 0.001),在第9周时处于低水平(p = 0.005)。血清IL - 5水平在第3周时升高(第0 - 3周;p = 0.01),在第9周时因反弹效应而降低(第6 - 9周,p = 0.003)。TNF - α水平随着开菲尔的饮用而升高(p = 0.046),但在配对比较中升高不显著,且在第0周和第6周之间处于临界水平(p = 0.013)。IL - 5和TNF - α水平在第9周时恢复到原始水平(第0周)。其他细胞因子(IL - 1和TGF - β)的水平在饮用开菲尔后未发生显著变化。这些结果表明,饮用开菲尔会增加免疫反应向TH1型极化,降低TH2型反应,从而减少过敏反应。饮用开菲尔导致的IL - 8水平降低可能通过抑制中性粒细胞趋化和活化来控制炎症反应。另一方面,也得出结论,IL - 5的增加可能会刺激胃肠道黏膜分泌IgA,从而在肠腔中产生更有效的免疫反应。