Analysis of Peripheral Blood Lymphocyte Subsets,NK Cells,and Delayed Type Hypersensitivity Skin Test in Patients With Premature Ovarian Failure

PROBLEM : Premature ovarian failure (POF) probably belongs to the group of autoimmune endocrinopathies. Cell-mediated immune parameters were investigated. Sex steroids have a profound effect on the immune system. POF patients and postmenopausal control women (PM) were tested with or without estrogen substitution. METHOD : A novel FACS analysis system (using double labeling techniques) was used in 30 patients with POF to enumerate the subjects of peripheral blood lymphocytes and NK cells. Eighteen PM women and 30 healthy men and women served as controls. We also tested the delayed type hypersensitivity skin test (DTH) toward Candida in the POF patient group to be informed on their cell-mediated immune function. RESULTS : The numbers of blood lymphocytes, CD3⁺, CD4⁺ and CD8⁺ T cells, were not abnormal in POF patients. However, HLA-DR⁺ T cells were increased in POF patients and in PM women (P<0.05). These elevated numbers were partially reversible by estrogen substitution. The number of CD19⁺ cells (B cells) was elevated, whereas CD3^?/CD16⁺/CD56⁺ cells (NK cells) were decreased in POF patients (P<0.05), irrespective of estrogen substitution. DTH skin tests toward 0.1% Candidin (0.1 ml intradermal injection) were negative in 11 out of 20 tested POF patients, compared to only 2 out of 10 tested controls (P<0.05). CONCLUSION : POF patients show numerous immune cell abnormalities. These abnormalities were only partially due to estrogen deficiency. We hypothesize that these abnormalities either lead to ovarian autoimmunity or may have direct effects on the ovarian function.     

Dietary supplementation of neutral and acidic oligosaccharides enhances Th1-dependent vaccination responses in mice

Immunomodulatory effects of oligosaccharide preparations that resemble chemical and functional aspects of human milk oligosaccharides (HMOS) were studied for the development of new concepts in infant nutrition. A dose range of 1-5% (w/w) dietary pectin-derived acidic oligosaccharides (AOS) was tested in a murine influenza vaccination model. In addition, combinations of AOS and a 9:1 mixture of galacto-oligosaccharides and long-chain fructo-oligosaccharides (GOS/FOS) were tested at a fixed total dietary dose of 2% (w/w). It was found that AOS significantly enhanced vaccine-specific delayed-type hypersensitivity (DTH) responses in a dose-dependent manner. This was accompanied by a reduction in T-helper2 (Th2) cytokine production by splenocytes in vitro. Overall, this indicates that the systemic immune response to the vaccine was Th1-skewed by the dietary intervention. Combinations of GOS/FOS and AOS were more effective in enhancing DTH responses than either of the oligosaccharides alone, suggesting interaction effects between these agents. Similar to effects in infants, supplementation of the murine diets with GOS/FOS and combinations of GOS/FOS and AOS for 6-wk enhanced the proportion of fecal bifidobacteria and lactobacilli, but AOS alone did not. In conclusion, these data indicate that GOS/FOS and AOS enhance systemic Th1-dependent immune responses in a murine vaccination model. As Th1-responses are weak in early life in humans, this might suggest that application of these oligosaccharides in infant formulas will be beneficial for the development of the infant's immune system.     

硒酵母的药理作用

选用多个正常或病理动物模型,观察了硒酵母(SEY)在免疫调节、保肝抗溃疡及抗心肌缺血等方面的作用.结果表明,SEY(30～90mg·kg~(-1)·d~(-1)×5～7d,ig)可上调迟发型变态反应小鼠低下的细胞免疫功能,并对四氯化碳引起的小鼠肝损伤与应激性小鼠胃出血均具有一定的保护作用.SEY(25～50mg·kg(-1)·d~(-1)×7d,ig)对垂体后叶素诱发的大鼠急性心肌缺血亦有一定的改善作用.提示SEY具有较广泛的药理作用.     

能量和锌对小鼠免疫功能的影响

实验研究观察了不同进食状况下小鼠的迟发型变态反应（ＤＴＨ）。用三种不同方法测定ＤＴＨ。结果表明：限制饮食可以抑制小鼠的ＤＴＨ，在正常进食下，适度加锌可增强小鼠的ＤＴＨ。     

小鼠自身免疫性睾丸炎 Ⅰ.精子特异性T细胞被动免疫模型

BALB/C小鼠的附睾精子经体外获能后作抗原,免疫同种小鼠,取引流淋巴结细胞培养,同时加入获能精子作第二次刺激,扩增后通过Percoll 密度梯度离心提取母化的淋巴细胞,在无抗原刺激下加白细胞介素—2培养,使已致敏的T 淋巴母细胞继续生长,而B 细胞因无刺激而被筛除。收集精子特异性T 淋巴母细胞经腹腔转移给同种雄性小鼠,1～3周后做睾丸病理检查,发现有严重的自身免疫性睾丸炎形成。病变是以淋巴细胞、巨噬细胞浸润伴生精上皮破坏为特征的迟发型变态反应(DTH)。实验结果说明精子致敏的T 细胞介导了抗原特异性自身免疫性睾丸炎。     

Induction of active immune suppression by co-immunization with DNA- and protein-based vaccines

Although immunization has been used for eliciting immune response, here we show that it can also induce immune suppression. When a DNA vaccine encoding a viral antigen such as the VP1 protein from the foot and mouth disease virus is administered together with its recombinant protein antigen or a viral preparation containing the same antigen, the immunized animals developed significantly reduced antigen-specific T cell-mediated responses and became impaired to subsequent rechallenge with the same antigen. The induction of immune suppression is mediated by suppressor T cells, as demonstrated by an adoptive transfer experiment and mixed lymphocyte reactions. The induction of immune suppression in immunized animals is also correlated with a shift of cytokine balance, as reflected by an elevated level of IL-10 and reduced level of IFN-gamma or IL-2. Hence, co-immunization with DNA- and protein-based vaccines may represent a novel means for inducing active suppression against untoward immunity.     

Skin tests predict survival after autologous tumor cell vaccination in metastatic melanoma: Experience in 81 patients

Background:Currently there is no standard adjuvant treatmentfollowing surgical resection of metastatic melanoma. We investigated whethersurgery followed by autologous tumor cell-BCG vaccination was beneficial formalignant melanoma patients. In this study we focus on the prognostic valueof DTH response following vaccination therapy. Patients and methods:Eighty-one patients with AJCC stage III andIV melanoma were selected. Whenever feasible, radical metastasectomy wasperformed. ASI was initiated by the administration of three weeklyintra-cutaneous vaccinations with 10⁷ irradiated autologous tumorcells, starting four weeks after surgery. Depending on the size of DTHresponse to the first three injections, subsequent vaccinations were planned.The first two vaccines also contained 10⁷ BCG organisms as animmune stimulatory adjuvant. Results:Induration as well as erythema correlated strongly withsurvival (P< 0.0001 and P= 0.0004). After radicalmetastasectomy in stage III melanoma patients a five-year survival of48% was observed. In stage IV disease, a five-year survival of34% was seen, after radical surgery had been performed. Whenmacroscopic disease was present at start of vaccination treatment, no clinicalresponses occurred. Apart from transient skin ulceration at the site ofBCG-containing vaccinations, no serious side effects were observed. Conclusions:This study shows that large-scale preparation ofautologous melanoma cell vaccines is feasible, while vaccination results inDTH responses that correlate significantly with survival. ASI seemed to bebeneficial in stage III and stage IV melanoma when given in the adjuvantsetting, while causing only very mild side effects.     

Characterization of a murine keyhole limpet hemocyanin (KLH)-delayed-type hypersensitivity (DTH) model: Role for p38 kinase

Molecular and cellular assessment of dermal delayed-type hypersensitivity (DTH) responses is a useful approach for evaluating the mechanism of action (MOA) of immunomodulatory agents. In the present report, we characterized the delayed-type hypersensitivity response induced by keyhole limpet hemocyanin (KLH), and validated its utility by evaluating an immunomodulator, BIRB-796. Intradermal KLH challenge of the ear pinna following subcutaneous antigen sensitization resulted in a pronounced skin inflammation that peaked at 24–48 h. At the molecular level, there was an activation of 3 mitogen-activated protein kinases (MAPKs: p38, JNK and ERK), an induction of the chemokines CCL2/JE, CXCL2/Mip-2, CXCL1/KC, CCL3/Mip-1α CCL4/Mip-1β and CXCL10/IP-10, and expression of the cytokines IL-1β and IL-10 in the ear parenchyma. Modulation of TNFα protein level was only detected in ex-vivo ear whole organ cultures (EWOC). Consistent with this inflammatory profile there was an infiltration of neutrophils and mononuclear cells into the ear parenchyma. BIRB-796, a potent allosteric p38 MAPK inhibitor attenuated the ear swelling response, which correlated with a reduced inflammatory profile. BIRB-796 inhibited p38 but not JNK or ERK kinase activation, decreased multiple chemokines which correlated with a decrease in the infiltration of neutrophils and macrophages; CD4 T cells were modesty reduced. Similarly, there was a decrease of levels of cytokines including IL-1β, IL-10 and TNFα. These data support the utility of this model for evaluating immunomodulators on skin inflammation and suggest that modulation of p38 kinase may be of therapeutic value for the treatment of inflammatory skin conditions.     

The in vivo and in vitro induction of anterior chamber associated immune deviation to myelin antigens in C57BL/6 mice

Introduction of antigens into the anterior chamber (AC) of the eye generates a specific systemic form of tolerance that is termed AC-associated immune deviation (ACAID). Experimental autoimmune encephalomyelitis (EAE) is an animal model of the human CNS demyelinating diseases, including multiple sclerosis (MS) and acute disseminated encephalomyelitis. We investigated whether the encephalitogenic antigens myelin oligodendrocyte glycoprotein (MOG_35–55) or myelin basic protein (MBP) induce ACAID in the EAE-prone C57BL/6 mice. We hypothesized that injection of MOG_35–55/MBP induces antigen-specific tolerance whether via the AC route, the adoptive transfer of in vitro-generated MOG_35–55-specific/MBP-specific ACAID antigen presenting cells (APCs), or the adoptive transfer of MOG_35–55-specific/MBP-specific ACAID T regulatory cells (Tregs). ACAID is characterized by the specific impairment of delayed-type hypersensitivity (DTH) responses. Thus, DTH assays were used to test for ACAID following the AC injection of MOG_35–55/MBP, or the intravenous injection of MOG_35–55-specific/MBP-specific ACAID APCs. The functional local adoptive transfer (LAT) assays were used to examine the putative regulatory functions of in vitro generated MOG_35–55-specific/MBP-specific Tregs. This report is the first to demonstrate the in vivo and in vitro induction of MOG_35–55-specific/MBP-specific ACAID-mediated tolerance in C57BL/6 mice. These findings highlight the need for novel immunotherapeutic strategies for MS and optic neuritis.