Rac1在小鼠中性粒细胞抗白假丝酵母菌感染中的作用

目的评价Rac1基因在小鼠抗白假丝酵母菌感染中的作用。方法 (1)β-actin作为内参,Western blot半定量法检测两种小鼠中性粒细胞Rac1蛋白表达量;(2)趋化实验用Zigmond法经甲酰甲硫氨酰-亮氨酰-苯丙氨酸(fMLP)诱导中性粒细胞移动,延时显微镜观察中性粒细胞移动并拍照,细胞追踪软件分析小鼠中性粒细胞趋化功能;(3)fMLP诱导小鼠中性粒细胞过氧化物的产生应用异氨基苯二酰肼化学发光分析法通过微孔板发光检测仪检测过氧化物酶含量,采用细胞色素c还原法分析豆蔻酸-佛波醇-乙酸酯(PMA)引发中性粒细胞产生的过氧化物。结果假丝酵母菌耐受BALB/c小鼠中性粒细胞Rac1蛋白表达量高于假丝酵母菌易感CBA/CaH小鼠;BALB/c小鼠中性粒细胞趋化运动功能较CBA/CaH小鼠强(P<0.05);经fMLP和PMA诱导后,BALB/c小鼠中性粒细胞过氧化物的产生量高于CBA/CaH小鼠(P<0.05)。结论在小鼠中性粒细胞抗白假丝酵母菌感染过程中,Rac1能增强小鼠中性粒细胞的杀伤能力。     

Racl在小鼠中性粒细胞抗白假丝酵母菌感染中的作用

目的 评价Racl基因在小鼠抗白假丝酵母菌感染中的作用.方法 (1)β-actin作为内参,Western blot半定量法检测两种小鼠中性粒细胞Rael蛋白表达量;(2)趋化实验用Zigmond法经甲酰甲硫氨酰-亮氨酰-苯丙氨酸(fMLP)诱导中性粒细胞移动,延时显微镜观察中性粒细胞移动并拍照,细胞追踪软件分析小鼠中性粒细胞趋化功能;(3)fMLP诱导小鼠中性粒细胞过氧化物的产生应用异氨基苯二酰肼化学发光分析法通过微孔板发光检测仪检测过氧化物酶含量,采用细胞色素c还原法分析豆蔻酸-佛波醇-乙酸酯(PMA)引发中性粒细胞产生的过氧化物.结果 假丝酵母菌耐受BALB/c小鼠中性粒细胞Racl蛋白表达量高于假丝酵母菌易感CBA/CaH小鼠:BALB/c小鼠中性粒细胞趋化运动功能较CBA/CaH小鼠强(P<0.05):经fMLP和PMA诱导后,BALB/c小鼠中性粒细胞过氧化物的产生量高于CBA/CaH小鼠(P<0.05).结论 在小鼠中性粒细胞抗白假丝酵母菌感染过程中,Racl能增强小鼠中性粒细胞的杀伤能力.     

牙龈卟啉单胞菌基因疫苗pcDNA3.1(+)/kgpcd免疫原性研究

目的:研究重组质粒pcDNA3. 1( ) /kgpcd免疫原性,并观察目的基因编码的蛋白在小鼠肌肉及颌下腺组织中的原位表达情况。方法:重组质粒pcDNA3. 1 ( ) /kgpcd经肌肉注射和颌下腺区注射免疫BALB/c小鼠,采用间接ELISA方法检测免疫后BALB/c小鼠血清中IgG和唾液中sIgA抗体水平;用免疫组织化学染色观察目的基因编码的蛋白在小鼠组织中的表达。结果:重组质粒经肌肉注射免疫可产生高水平的特异性血清IgG抗体,颌下腺区注射可同时诱导高水平的唾液中sIgA和血清中IgG抗体;骨骼肌、颌下腺导管内、腺泡腔内可见阳性着色。结论:重组质粒pcDNA3. 1( ) /kgpcd经颌下腺注射可同时诱导黏膜免疫和全身免疫应答,可作为有效免疫原,这为免疫牙周炎模型定菌鼠实验提供了依据。     

68株临床分离口腔念珠菌对4种抗真菌药的体外药敏实验结果分析

目的：评价临床致病性口腔念珠菌感染的种类及对氟康唑、两性霉素B、5-氟胞嘧啶和伊曲康唑的药物敏感性,比较各种口腔黏膜病中的感染菌株的种类和耐药比例,为临床用药提供指导。方法：收集临床菌株,经YBC Test Kit鉴定其念珠菌种类;根据NCCLS的M27-A2标准方案测定3组（口腔念珠菌病、口腔扁平苔藓合并口腔念珠菌感染、头颈部放化疗患者合并口腔念珠菌感染）菌株对4种抗真菌药物的最低抑菌浓度,并比较不同口腔黏膜病合并念珠菌感染菌株的耐药状况。结果：68株临床分离株对5-氟胞嘧啶全部敏感,91．2％对两性霉素B敏感,氟康唑和伊曲康唑的耐药比例分别为13．2％和22．1％,放化疗组的非白色念株菌比例及耐药比例明显高于口腔念珠菌病组和扁平苔藓组。结论：口腔念珠菌感染菌株对氟康唑和伊曲康唑存在较高耐药比例和交叉耐药比例,头颈部肿瘤放疗或化疗患者口腔念珠菌感染菌株的非白色念株菌比例及耐药比例明显高于口腔念珠菌病和扁平苔藓合并口腔念珠菌感染菌株。     

HIV感染患者口腔念珠菌的培养鉴定及耐药性研究

目的：研究HIV感染患者口腔念珠菌感染的菌株种类和耐药情况,比较CHROMagar显色培养基（CHROMagar）和API 20C AUX酵母菌鉴定系统（API）在HIV感染者口腔念珠菌鉴别中的应用,指导临床抗真菌药物的使用。方法：6株念珠菌标准菌株和17例HIV感染合并口腔念珠菌病患者舌背白色刮取物,分别接种于沙氏培养基和CHROMagar,37℃48h后取沙氏培养基上生长物作API 20C AUX鉴定和耐药性检测。结果：白色念珠菌8例,热带念珠菌4例,近平滑念珠菌4例,白色念珠菌和克柔念珠菌混合感染1例。念珠菌对特比萘芬耐药,对氟康唑、酮康唑、眯康唑、克霉唑中度敏感,对制霉菌素、伊曲康唑、两性霉素B、氟胞嘧啶高度敏感。API 20C AUX、CHROMagar在鉴定白色念珠菌上无显著性差异,API20CAUX鉴定非白色念珠菌准确率高于CHROMagar,但CHROMagar可鉴别混合菌种感染。结论：HIV感染患者口腔念珠菌以非白色念珠菌居多（52．9％）,分型初筛可用CHROMagar,准确分型需用API20CAUX鉴定,两种方法互有裨益。临床用药建议选择制霉菌素、伊曲康唑、两性霉素B、氟胞嘧啶中的两种任意综合治疗。     

分泌特异性牙龈拟杆菌单克隆抗体杂交瘤的建立

牙龈拟杆菌Bg在产黑色素拟杆菌群中毒力最强,现已公认Bg是牙周炎、急性周尖炎的关键细菌。本实验用Bg 47A—1 整体细菌免疫Balb/c小鼠,脾细胞与SP2/0小鼠骨髓瘤细胞融合,通过筛选、克隆化。获得了一株能够整定分泌Bg单克隆抗体杂交瘤系。特异性鉴定表明该单克隆抗体不与所试验的5类20株产黑色素拟杆菌群菌株(非Bg菌)和其它口腔常见菌(变形链球菌、巨核梭形杆菌、脆弱拟杆菌)发生交叉反应。免疫扩散法证实比单抗属于IgG2b。所制备腹水IgG含量高,抗体效价1:50000以上,预期可用于临床标本中Bg的鉴定与栓出及组织中定位研究。     

抗粘放菌5519Ⅰ型菌毛单克隆抗体的制备和鉴定

目的制备抗粘放菌5519Ⅰ型菌毛单克隆抗体。方法采用BALB/C小鼠免疫后取脾细胞与SP2/0小鼠骨髓瘤细胞杂交融合,制备单克隆抗体,并以ELISA法和双向琼脂扩散法进行鉴定。结果共得到6株杂交瘤细胞,抗体效价达110～100万,类型为IgG,亚类为IgG1。结论经BALB/C小鼠免疫可得到高纯度的抗粘放菌Ⅰ型菌毛单克隆抗体,而且效价高,细胞株稳定分泌抗体。     

维生素A对牙周致病菌免疫小鼠免疫应答调节的作用

目的:探讨维生素A缺乏对特异性牙周致病菌-伴放线放线杆菌(A.actinomycetetemcomitans,Aa)引起的小鼠免疫应答作用的影响.方法:整个实验过程采用无维生素A饮食(vitamine A-depleted diet,VAD)或常规维生素A饮食(vitamine A-sufficient regular diet,RD)喂养BALB/c鼠.2周后,免疫Aa建立免疫动物模型,6周后处死小鼠,ELISA法测定血清中的抗Aa特异性抗体总IgG、IgM及IgG亚类抗体滴度及细胞上清中细胞因子的浓度,3H-Tdr掺入法测定T细胞增殖反应.实验结果采用SPSS 11.5软件包进行统计学分析.结果:Aa免疫的总IgG和IgM抗体水平明显升高,非免疫组则不能产生抗体.Aa免疫+VAD组与Aa免疫+RD组相比,总IgG水平显著升高(P<0.05);IgG2a的抗体水平明显增加,而IgG1亚型的抗体水平却明显降低,差异显著(P<0.05).Aa免疫组可诱导机体产生较强的特异性T细胞免疫反应,而Aa免疫+VAD组T细胞增殖反应明显高于Aa免疫+RD组,具有统计学差异(P<0.05);细胞上清中RANKL、IFN-γ及TNF-α的表达增加,IL-10的表达降低(P<0.05).结论:饮食中维生素A缺乏,可增加Aa免疫鼠引起的免疫炎症反应,提示充足的维生素A是维持机体健康的重要因素.     

口腔粘膜白色念珠菌感染的动物模型研究

口腔色念珠菌感染的动物模型虽有报道，但成功率不高。本研究通过：（１）用从患者口腔中分离的白色念珠菌株并通过兔全身感染再分离的方法，加强菌株的毒性；（２）扩大口腔检查部位的范围，成功地（１００％）在大白鼠口腔中产生白色念珠菌感染，并对其是否为口腔粘膜癌前病变进行讨论。     

白念珠菌口腔黏膜感染小鼠模型的建立及评估

［摘要］ 目的 建立白念珠菌口腔黏膜感染ICR小鼠模型,动态观察口腔黏膜及相应的全身感染情况,并对变化情况进行初步分析。方法 将白念珠菌接种于免疫功能低下小鼠口腔,采用肉眼观察口腔舌背病损改变、口腔内白念珠菌菌量检测、组织病理学等方法评估口腔内黏膜感染情况,检测体重、载菌量（肾、肝、下降结肠内粪便）评估全身感染情况。观察1周,并记录数据。空白对照组为接种生理盐水的免疫功能低下组。结果 ①小鼠接种白念珠菌后第1天口腔白念珠菌菌量较低,随后迅速增长,3~7 d内趋于稳定,稳定于106~107CFU/mL。同时口腔舌背病损也出现类似趋势,第1天未见明显伪膜,3~7 d内可见舌背伪膜存在。②病理切片PAS染色显示接种后3~5 d内白念珠菌形成菌丝侵入并破坏上皮,到接种第7天时,黏膜上皮多见酵母细胞。③小鼠接种白念珠菌后体重逐渐下降。粪便载菌量逐渐上升,第5天达到最大,但肾、肝无白念珠菌感染。空白组未见白念珠菌感染。结论 通过在ICR小鼠口腔内接种白念珠菌可以建立白念珠菌口腔黏膜感染动物模型。接种后白念珠菌感染程度在1周内存在变化。动物实验应根据感染状况选择合适的研究时段。     

Active and passive immunization against oral Candida albicans infection in a murine model

BACKGROUND/AIMS: Clinical and laboratory studies are consistent with a major role for cell-mediated immunity in recovery from oral infection with Candida albicans, but the role of humoral immunity remains controversial. The purpose of this study was to establish the relative contributions of cellular and humoral immunity to protection against oral candidiasis in a murine model, and to determine whether host responses could be enhanced by different immunization strategies. RESULTS: Active oral immunization was protective in BALB/c and CBA/CaH mice, reducing both fungal burden and duration of infection after secondary challenge, whereas systemic immunization failed to protect against subsequent oral challenge. Candida-specific IgM was the predominant antibody detected in serum following both primary and secondary oral challenge; however, Candida-specific salivary IgA was not detectable. Immunization by passive transfer of either lymphocytes or immune serum did not confer any significant protection against oral infection in either susceptible or resistant mouse strain. CONCLUSION: The data demonstrate a possible role for mucosa-associated immunity following active immunization by the oral route, most likely exerted by local T lymphocytes resident in the oral mucosa, but there was no evidence to support a role for humoral immunity in protection against oral candidiasis.     

Differences in mouse strain influence leukocyte and immunoglobulin phenotype response to Porphyromonas gingivalis

This study examined the nature of the infiltrating cells in Porphyromonas gingivalis-induced lesions and immunoglobulins in the serum samples of BALB/c (H-2d), C57BL6 (H-2b), DBA/2J (H-2d) and CBA/CaH (H-2k) mice. Mice were immunized intraperitoneally with P. gingivalis outer membrane antigens or sham-immunized with phosphate-buffered saline followed by subcutaneous challenge with live organisms 1 week after the final immunization. The resulting skin abscesses were excised 7 days later, cryostat sections cut and an immunoperoxidase method used to detect the presence of CD4+ and CD8+ T-cell subsets, CD14+ macrophages and CD19+ B cells. Peroxidase positive neutrophils and IgG1- and IgG2a-producing plasma cells were also identified. Anti P. gingivalis IgG1 and IgG2a subclass antibodies were determined in serum obtained by cardiac puncture. Very few CD8+ T cells and CD19+ B cells were found in any of the lesions. The percentages of CD4+ cells, CD14+ cells and neutrophils were similar in lesions of immunized BALB/c and C57BL6 mice, with a trend towards a higher percentage of CD14+ cells in sham-immunized mice. The percentage of CD14+ cells was higher than that of CD4+ cells in immunized compared with sham-immunized DBA/2J mice. The percentages of CD4+ and CD14+ cells predominated in immunized CBA/CaH mice and CD4+ cells in sham-immunized CBA/CaH mice. The percentage of neutrophils in immunized CBA/CaH mice was significantly lower than that of CD14+ cells and CD4+ cells in sham-immunized mice. IgG1+ plasma cells were more dominant than IgG2a+ cells in immunized BALB/c, C57BL6 and DBA/2J mice, whereas IgG2a+ plasma cells were more obvious in sham-immunized mice. IgG2a+ plasma cells were predominant in immunized and sham-immunized CBA/CaH mice. In the serum, specific anti-P. gingivalis IgG2a antibody levels (Th1 response) were higher than IgG1 levels (Th2 response) in sham-immunized CBA/CaH and DBA/2J mice. In immunized BALB/c mice, IgG2a levels were lower than IgG1 levels, while IgG2a levels were higher in immunized C57BL6 mice. In conclusion, this study has shown differences in the proportion of infiltrating leukocytes and in the subclasses of immunoglobulin produced locally and systemically in response to P. gingivalis in different strains of mice, suggesting a degree of genetic control over the response to P. gingivalis.     

Genetic variation in the recognition of Porphyromonas gingivalis antigens in mice

T-cell cytokine profiles, anti-Porphyromonas gingivalis antibodies and Western blot analysis of antibody responses were examined in BALB/c, CBA/CaH, C57BL6 and DBA/2J mice immunized intraperitoneally with different doses of P. gingivalis outer membrane antigens. Splenic CD4 and CD8 cells were examined for intracytoplasmic interleukin (IL)-4, interferon (IFN)-gamma and IL-10 by FACS analysis and levels of anti-P. gingivalis antibodies in the serum samples determined by enzyme-linked immunosorbent assay. Western blot analysis was performed on the sera from mice immunized with 100 microg of P. gingivalis antigens. The four strains of mice demonstrated varying degrees of T-cell immunity, although the T-cell cytokine profiles exhibited by each strain were not affected by different immunizing doses. While BALB/c and DBA/2J mice exhibited responses that peaked at immunizing doses of 100-200 microg of P. gingivalis antigens, CBA/CaH and C57BL6 demonstrated weak T-cell responsiveness compared with control mice. Like the T-cell responses, serum antibody levels were not dose dependent. DBA/2J exhibited the lowest levels of anti-P. gingivalis antibodies followed by BALB/c with CBA/CaH and C57BL6 mice demonstrating the highest levels. Western blot analysis showed that there were differences in reactivity between the strains to a group of 13 antigens ranging in molecular weight from 15 to 43 kDa. Antibody responses to a number of these bands in BALB/c mice were of low density, whereas CBA/CaH and C57BL6 mice demonstrated high-density bands and DBA/2J mice showed medium to high responses. In conclusion, different immunizing doses of P. gingivalis outer membrane antigens had little effect on the T-cell cytokine responses and serum anti-P. gingivalis antibody levels. Western blot analysis, however, indicated that the four strains of mice exhibited different reactivity to some lower-molecular-weight antigens. Future studies are required to determine the significance of these differences, which may affect the outcome of P. gingivalis infection.     

Differences in mouse strain influence leukocyte and immunoglobulin phenotype response to Porphyromonas gingivalis

This study examined the nature of the infiltrating cells in Porphyromonas gingivalis-induced lesions and immunoglobulins in the serum samples of BALB/c (H-2^d), C57BL6 (H-2^b), DBA/2J (H-2^d) and CBA/CaH (H-2^k) mice. Mice were immunized intraperitoneally with P. gingivalis outer membrane antigens or sham-immunized with phosphate-buffered saline followed by subcutaneous challenge with live organisms 1 week after the final immunization. The resulting skin abscesses were excised 7 days later, cryostat sections cut and an immunoperoxidase method used to detect the presence of CD4⁺ and CD8⁺ T-cell subsets, CD14⁺ macrophages and CD19⁺ B cells. Peroxidase positive neutrophils and IgG1- and IgG2a-producing plasma cells were also identified. Anti P. gingivalis IgG1 and IgG2a subclass antibodies were determined in serum obtained by cardiac puncture. Very few CD8⁺ T cells and CD19⁺ B cells were found in any of the lesions. The percentages of CD4⁺ cells, CD14⁺ cells and neutrophils were similar in lesions of immunized BALB/c and C57BL6 mice, with a trend towards a higher percentage of CD14⁺ cells in sham-immunized mice. The percentage of CD14⁺ cells was higher than that of CD4⁺ cells in immunized compared with sham-immunized DBA/2J mice. The percentages of CD4⁺ and CD14⁺ cells predominated in immunized CBA/CaH mice and CD4⁺ cells in sham-immunized CBA/CaH mice. The percentage of neutrophils in immunized CBA/CaH mice was significantly lower than that of CD14⁺ cells and CD4⁺ cells in sham-immunized mice. IgG1⁺ plasma cells were more dominant than IgG2a⁺ cells in immunized BALB/c, C57BL6 and DBA/2J mice, whereas IgG2a⁺ plasma cells were more obvious in sham-immunized mice. IgG2a⁺ plasma cells were predominant in immunized and sham-immunized CBA/CaH mice. In the serum, specific anti-P. gingivalis IgG2a antibody levels (Th1 response) were higher than IgG1 levels (Th2 response) in sham-immunized CBA/CaH and DBA/2J mice. In immunized BALB/c mice, IgG2a levels were lower than IgG1 levels, while IgG2a levels were higher in immunized C57BL6 mice. In conclusion, this study has shown differences in the proportion of infiltrating leukocytes and in the subclasses of immunoglobulin produced locally and systemically in response to P. gingivalis in different strains of mice, suggesting a degree of genetic control over the response to P. gingivalis.     

The influence of genetic variation on the splenic T cell cytokine and specific serum antibody responses to Porphyromonas gingivalis in mice

BACKGROUND: T cell cytokine profiles in the spleens and anti-Porphyromonas gingivalis antibodies in the sera of P. gingivalis-immunized BALB/c (H-2d), CBA/CaH (H-2k), C57BL6 (H-2b), and DBA/2J (H-2d, C5 deficient) mice were examined. METHODS: Mice were immunized either by intraperitoneal injections of P. gingivalis outer membrane antigens and Freund's incomplete adjuvant weekly for 3 weeks or sham-immunized with PBS and adjuvant, followed by subcutaneous challenge with live organisms 1 week after the final immunization. Spleens were excised and blood samples collected by heart puncture at 0 and 7 days after challenge. Splenic CD4 and CD8 cells were stained for intracytoplasmic interleukin (IL)-4, interferon (IF)-gamma, and IL-10 and levels of anti-P. gingivalis antibodies in the serum samples determined by ELISA. RESULTS: Lesion sizes in immunized BALB/c mice remained stable for the 7-day experimental period. Immunized CBA/CaH and C57BL6 mice exhibited large lesions at day 1 reducing by day 7 particularly in the latter strain. Lesions in immunized DBA/2J mice were still larger than the other strains at day 7. With the exception of DBA/2J mice, sham-immunized mice demonstrated lesions which did not show signs of healing by day 7. T cell cytokine responses in sham-immunized mice at day 0 were low, increasing to a variable degree by day 7 after challenge in the 4 strains. Immunized BALB/c mice demonstrated intermediate T cell responses while generally exhibiting a stronger IFN-gamma response than IL-4 or IL-10. Immunized CBA/CaH and C57BL6 mice showed weak T cell cytokine responses while immunized DBA/2J displayed the strongest T cell responses particularly in regard to IL-4 positive cells. Sham-immunized mice had low levels of serum anti-P. gingivalis antibody levels at day 0 with levels increasing significantly by day 7 after challenge. Antibody levels in immunized mice seemed to correlate with lesion sizes. Immunized C57BL6 mice had the highest antibody levels followed by CBA/CaH, BALB/c with DBA/2J exhibiting low levels. The T cell and B cell antibody responses in each strain appeared to exhibit an inverse relationship. CONCLUSIONS: This study has shown that genetic differences at the level of H-2 haplotype induce variations in the local and T and B cell responses to P. gingivalis antigens. The responses of DBA/2J mice which have the same haplotype as BALB/c mice suggest that factors other than H-2 haplotype such as the C5 deficiency may influence this immune response. The significance of the specific antibody and T cell responses and of their inverse relationship to susceptibility to periodontal disease remains to be determined.     

Irradiation-induced oral candidiasis in an experimental murine model

The aim of this experiment was to establish a mouse model of irradiation-induced oral candidiasis and to explore the cellular populations and mechanisms by which the infection is cleared from the oral mucosa. BALB/c mice received irradiation to the head and neck equivalent to 800 Rad using a Cobalt 60 gamma source. Both irradiated and non-irradiated mice were infected orally with 1 x 10(8) Candida albicans yeasts. Compared with untreated controls, irradiated animals developed a more severe infection of longer duration, with hyphae penetrating the oral mucosa. Monoclonal antibody depletion of CD4+ but not CD8+ T cells from the systemic circulation prolonged the infection in irradiated mice, but not in controls. Supernatants of submandibular and superficial cervical lymph node cultures from irradiated animals demonstrated significantly higher titers of interleukin-12, but similar levels of interferon-gamma compared with controls. Screening for cytokine production by an RNase protection assay detected only macrophage migration inhibition factor in irradiated and non-irradiated oral tissues from day 8 onwards. The results of this study demonstrate a requirement for CD4+ T cells in the recovery from oral candidiasis induced by head and neck irradiation in mice, and are consistent with a role for Th1-type cytokines in host resistance.     

Gene targeting demonstrates that inducible nitric oxide synthase is not essential for resistance to oral candidiasis in mice, or for killing of Candida albicans by macrophages in vitro

Introduction:  Oral candidiasis is caused by opportunistic infections with the yeast Candida albicans . Previous studies have demonstrated important roles for innate immunity and T helper type 1-mediated inflammatory reactions in recovery from infection, with macrophages and neutrophils as key effector cells. Both effector cell types use the inducible isoform of nitric oxide synthase (iNOS) to generate candidacidal molecules, but it is not clear whether nitric oxide (NO) is an absolute requirement for candidacidal effector activity.
Methods:  In this study we directly investigated the role of iNOS-derived NO in resistance to murine experimental oral candidiasis, using iNOS knockout mice.
Results:  Knockout mice were no more susceptible to oral candidiasis than wild-type controls. Bone marrow-derived macrophages from the knockout mice killed C. albicans yeasts efficiently in vitro , and were still able to produce nitrites in an iNOS-independent manner, albeit less efficiently than wild-type controls. There were no significant differences in local mucosal production of interleukins 6, 12, 17A, or 23, interferon-γ, or transforming growth factor-β 24 h after oral challenge with C. albicans .
Conclusion:  These data suggest that iNOS-derived NO is not required for resistance to oral candidiasis in vivo , and that bone marrow-derived macrophages may have iNOS-independent means of generating reactive nitrogen species.     

Studies on the induction of the humoral immune responses to Bacteroides gingivalis fimbrial antigen in mice

Serum and salivary antibody responses to Bacteroides gingivalis fimbriae administered either orally or subcutaneously (s.c.) with or without an adjuvant in various strains of mice were examined in this study. Following results were obtained. 1) Oral administration of B. gingivalis fimbriae with GM-53 as an adjuvant in liposomes, but not in Tris-HCl buffer, definitely enhanced the fimbriae-specific IgG responses, mainly IgG1 followed by IgG2b, IgG2a and IgG3 in serum and IgA response in saliva of BALB/c mice. On the other hand, s.c. injection of fimbriae with GM-53 or MDP-Lys (L18) also raised the fimbriae-specific IgG followed by IgA and IgM responses in serum, and both IgA and IgG responses in saliva of BALB/c mice. Oral immunization was less effective than s.c. injection in terms of the production of serum antibody in the mice. However, the level of salivary antibody of mice injected s.c. was similar to that of mice immunized orally. 2) High anti-fimbriae antibodies in serum were maintained in BALB/c mice immunized orally with fimbriae and GM-53 in liposomes for approximately 7 months after the primary immunizations. Oral administration also induced and held the fimbriae-specific IgA response in saliva for at least 6 months after the primary immunizations. The levels of fimbriae-specific IgA in saliva after the second boosters on days 123 and 124 were higher than those after the primary ones on days 27 and 28. 3) Among various strains of mice immunized orally with fimbriae and GM-53 in liposomes, BALB/c and DBA/2 mice (H-2d) significantly produced high levels of both serum IgG and salivary IgA antibodies specific for fimbriae. Furthermore, B10.D2 mice (H-2d) were responders followed by B10.BR (H-2k), while C57BL/10 mice (B10, H-2b) were low responders to the fimbriae. These results show that the combined use of fimbriae together with an adjuvant results in a sharply increased IgA antibody response in saliva and a predominantly stimulated IgG antibody in serum, and it was suggested that these responses are restricted by H-2 haplotype.