结核菌素试验复强反应与血清IgE含量关系的研究

报告一处农村全人口结核菌素试验复强反应现象调查。复强反应阳性率为１１．０％，采用ＥＬＩＳＡ测定ＰＰＤ试验前后５组人群血清ＩｇＥ含量，结果：人群血清ＩｇＥ含量随着ＰＰＤ试验次数的增加呈现逐渐升高的趋势。     

血清抗结核菌纯蛋白衍生物抗体测定的价值

结核感染是肾移植术后的常见并发症，且早期不易诊断。血清抗结核菌纯蛋白衍生物抗体（抗ＰＰＤ－ＩｇＧ）检测的结果表明，并发活动性结核的病人血清抗ＰＰＤ－ＩｇＧ均呈阳性反应，据此诊断并发结核的平均时间为２０．５天，明显早于根据细菌学、病理学或影像学确诊的平均时间（６６．８天）。肾移植后因其他感染或排斥反应引起的发热，抗ＰＰＤ－ＩｇＧ水平则无明显增高；经治疗结核感染得到控制的病人，血清抗ＰＰＤ－ＩｇＧ亦恢复正常。由此表明，血清特异性抗体检测是早期诊断肾移植后并发结核感染的良好指标。     

结核菌感染者微卡菌苗预防作用的研究

目的 观察、评价母牛分枝杆菌菌苗（微卡苗）对结核菌感染健康人群ＰＰＤ皮试强阳性者预防性治疗的效果。方法 对１８～４５岁ＰＰＤ皮试强阳性健康人群分为微卡苗注射组、口服ＩＮＨ组和未处理对照组,观察对比治疗前、后的ＰＰＤ皮试反应变化及发病情况。结果 治疗后较治疗前ＰＰＤ反应：微卡苗组硬结平均直径显著减小（Ｐ〈０．０１）,出现水泡、坏死者显著减少（Ｐ〈０．０１）,ＩＮＨ预防组和未处理对照组二者变化较小（Ｐ     

慢性肾功能衰竭接受替代治疗病人并发结核病

为了解慢性肾功能衰竭治疗中并发结核情况，对我院近５年收治的９７８例慢性肾功能衰竭接受替代治疗的病人进行了分析，其中４０例并发结核病，发生率为４．１％，为一般人群的１０倍。结核病６８％发生于透析或肾移植术后１年之内。其中肺外结核占８２．５％，淋巴结核是最常见的表现类型。慢性肾功能衰竭并发结核病的诊断仍有一定困难，结核菌素皮试在这类病人的阳性率仅６．４％。检测血清抗结核菌纯蛋白衍生物抗体（ＰＰＤ－ＩｇＧ）是一个有价值的辅助诊断方法，本组活动性结核８５％（３４／４０）呈阳性反应。并发浆膜腔或泌尿系结核的病人，检测渗出液或尿的抗ＰＰＤ－ＩｇＧ，或以聚合酶链反应（ＰＣＲ）检测结核菌ＤＮＡ有助于诊断。     

ELISA法检测胸液抗PPD－IgG对结核性和恶性胸液的鉴别意义

应用酶联免疫吸附分析法（ＥＬＩＳＡ）检测胸液抗人型结核菌ＰＰＤ抗体ＩｇＧ水平，对结核性胸膜炎诊断的灵敏性８９．０％，特异性９１．５％。提示此法可为与恶性胸腔积液鉴别诊断的一种辅助检查。     

亲和层析纯化胰吸虫抗原免疫活性的鉴定

本文采用对流免疫电泳、ＳＤＳ－ＰＡＧＥ和免疫印渍技术对亲和层析纯化胰吸虫抗原（ＥＰＰ）的免疫活性作了初步鉴定。结果显示：ＥＰＰ与兔免疫血清在对流免疫电泳中出现沉淀线，ＥＰＰ经ＳＤＳ－ＰＡＧＥ、考马斯亮蓝Ｒ－２５０染色后显示４条蛋白多肽，分子量约为５９～６２、４９、２７～２９、１５ＫＤ；转印至ＮＣ膜上的ＥＰＰ与兔免疫血清呈现２条带，分子量为４９、２７ＫＤ，而与胰吸虫病牛血清则主要出现４９ＫＤ的反应带：揭示该亲和层析纯化抗原具有免疫诊断价值。     

ABC－ELISA法检测脑脊液抗PPD－IgG及其临床意义

用ＡＢＣ－ＥＬＩＳＡ法检测５５例结脑和３７例非结脑患者脑脊液抗ＰＰＤ－ＩｇＧ并与ＥＬＩＳＡ法比较。结果两法阳性率分别为９６．４％和８３．６％，ＡＢＣ－ＥＬＩＳＡ法的敏感性高于ＥＬＩＳＡ法，Ｐ＜０．０５，而两法特异性均为９７．３％。同时检测结脑患者脑脊液和血清抗ＰＰＤ－ＩｇＧ，脑脊液阳性率为９６．４％高于血清阳性率为８５．５％，Ｐ＜０．０５。结果表明本法可为结脑诊断的一种辅助方法。     

血清结核抗体检测及结核菌素试验对肺结核病诊断的价值

目的 评价血清抗ＰＰＤ－ＩｇＧ检测及ＰＰＤ皮试在肺结核病诊断中的价值。方法 对１６２例肺结核病患者，４０例非结核病患者的ＰＰＤ皮试结果及血清抗ＰＰＤ－ＩｇＧ检测结果进行相关回归分析及ｘ＾２检验。结果 ＰＰＤ皮试敏感度为７２．８％，特异度为６０％；血清抗ＰＰＤ－ＩｇＧ检测敏感度为７９％，特异度为５５％，两者联合检测敏感度８９．５％；双阳性的特异度８５％；双阴性排除肺结核的特异度９７％。有无空洞对两者     

ABC—ELISA法检测脑脊液抗PPD—IgG及其临床意义

用ＡＢＣ－ＥＬＩＳＡ法检测５５例结脑和３７例非结脑患者脑脊液抗ＰＰＤ－ＩｇＧ并与ＥＬＩＳＡ法比较。结果两法阳性率分别为９６．４％和８３．６％，ＡＢＣ－ＥＬＩＳＡ法的敏感性高于ＥＬＩＳＡ法，Ｐ＜０．０５，而两法特异性均为９７．３％。同时检测结脑患者脑脊液和血清抗ＰＰＤ－ＩｇＧ，脑脊液阳性率为９６．４％高于血清阳性率为８５．５。结果表明本法可为结脑诊断的一种辅助方法。     

5种方法联合检测力阴肺结核诊断价值的研究

目的 探讨痰聚合酶链反应（ＰＣＲ）ＴＢ－ＤＮＡ检测、血清结核分支杆菌糖脂免疫球蛋白Ｇ（ＬＡＭＩｇＧ）、卡介菌免疫球蛋白Ｇ（ＰＰＤＩｇＧ）、结核特异性循环免疫复合物（ＳＣＩＣ）与结核菌素（ＰＰＤ）０．１Ｕ皮试联合检测对菌阴肺结核的诊断价值。方法 以上述５种检测方法用于力阳肺结核３１例、健康对照５３例、非结核肺疾病３０例、初治菌阴肺结核５４例同步检测。血清免疫学检测用酶联免疫吸附试验（ＥＬＩＳＡ）。对     

Serodiagnosis of tuberculosis by radioimmunoassay

Mycobacteria antigens derived from whole cells and cell walls of M. tuberculosis and M. bovis (BCG) and soluble purified protein derivative (PPD) prepared from M. tuberculosis were used in solid phase radioimmunoassays to measure the amount of reactive IgG antibody in serums from 54 patients with active (culture-positive) tuberculosis (Group I), 6 patients with inactive (culture-negative) tuberculosis (Group II), 15 healthy subjects who were skin test positive to PPD (Group III), and 30 healthy persons who were PPD skin test negative (Group IV). Patients with active tuberculosis had statistically larger (p less than 0.001) amounts of IgG antibody to M. tuberculosis whole cells, cell walls, and PPD and to BCG whole cells and cell walls when compared with the amount of antibody in serums from healthy subjects who were PPD skin test negative. However, no significant differences were detected in the mean antibody response or frequency of positive antibody responses between patients with active disease and those in clinical remission. Moreover, significant amounts of antibody were detected in 7 to 20% of healthy, tuberculin-reactive subjects. On the basis of these results, it is unlikely that antibody assays alone will prove useful in the diagnosis of this disease.     

The effect of exposure of hospital employees to patients with tuberculosis on dermal reactivity to four new tuberculins

An early (6-8 h) erythematous response to Purified Protein Derivative and to sonicate antigens (new tuberculins) prepared from Mycobacterium tuberculosis, M. vaccae, M. scrofulaceum, and M. leprae occurred much more frequently amongst hospital employees exposed to patients with tuberculosis than amongst factory workers. Biopsies taken from the skin test sites at 48 h revealed a more intense inflammatory cell infiltrate in response to PPD and the sonicate of M. tuberculosis, but not to the antigens of the other mycobacteria, amongst the hospital employees thus indicating a degree of specificity. The early response appears to be directed towards species specific antigens, but not, apparently, to the same as those that elicit the 48 h reactions. The hospital employees also had higher peripheral blood B-cell counts and total IgG levels, suggestive of an adjuvant effect. It is postulated that the early reaction results from repeated exposure to tubercle bacilli and the possible nature of the reaction is discussed.     

Evaluation of DPPD, a single recombinant Mycobacterium tuberculosis protein as an alternative antigen for the Mantoux test.

Although the tuberculin test has aided in the diagnosis of tuberculosis for more than 85 years, its interpretation is difficult particularly because sensitization with non-tuberculous mycobacteria leads to false positive tests. Using the guinea pig model of tuberculosis, we have recently described a recombinant antigen (DPPD) that could circumvent this problem. The DPPD gene is unique to the M. tuberculosis complex organisms and is absent in the organisms representative of all other members of the Mycobacterium genus. Moreover, DPPD induced strong DTH in 100% of the guinea pigs infected with M. tuberculosis and in none of the guinea pigs immunized with nine different species of Mycobacterium. Here we present results of a clinical investigation using DPPD. Mantoux test using both PPD and DPPD was initially performed in 26 patients with confirmed pulmonary tuberculosis and in 25 healthy PPD negative individuals. The results indicated that both PPD and DPPD elicited DTH in 24 out of the 26 patients. No DTH was observed in any of the PPD negative individuals. In addition, a small clinical trial was performed in a population of 270 clinically healthy and randomly selected individuals. DPPD produced a bimodal histogram of skin reaction size and PPD produced a skewed histogram. Because the DPPD gene is not present in non-tuberculous bacilli, these results suggest that this molecule can be an additional tool for a more specific diagnosis of tuberculosis.     

Clinical immunology of tuberculosis

The standard tuberculin skin test has been known as the prototype of delayed type hypersensitivity testing which is mediated by T cells and macrophages and plays an important role in the pathogenesis of tuberculosis. Tuberculosis is indeed a chronic infectious disease, but variation in the host immune responses to tubercle bacilli results in the various clinical manifestations of the disease ranging from an immunologically hyperreactive state observed in pleural fluid lymphocytes in tuberculous pleurisy to an almost totally unresponsive state observed in those severely ill with refractory tuberculosis. In tuberculous pleurisy, T cells in pleural fluid respond remarkably in vitro to PPD tuberculin whereas T cells in peripheral blood responded poorly to PPD stimulation. Compartmentalization of PPD-reactive T cells in the pleural fluid and immunosuppression by T cells and/or macrophages in the peripheral blood were responsible for this immunological difference observed between the lymphocytes in pleural fluid and those in peripheral blood of tuberculous pleurisy. In advanced, drug-resistant tuberculosis as well as in nontuberculous mycobacterial infection, the proliferative responses of T cells in vitro to PPD stimulation were impaired. This depressed T cell response was due to depressed interleukin-2 (IL-2) production and not due to depressed IL-2 responsiveness. Therefore, the addition of exogenous IL-2, returned the depressed PPD-induced lymphocyte proliferation in vitro in these patients to the level of the response observed in lymphocytes from patients with newly-diagnosed tuberculosis. Our results suggest that recombinant IL-2 offers a novel approach to the therapy of advanced, drug-resistant tuberculosis and nontuberculous mycobacterial infection. Preliminary clinical trials of immunotherapy with recombinant IL-2 reveals the effectiveness of this therapy and encourages us to extend the trial to a larger scale. Tubercle bacilli have various biological activities. Research on tuberculosis and tubercle bacilli have contributed much to the progress of biochemistry, pathology and immunology. Mycobacterium is a fascinating organism, which now presents another big appeal to those studying immunology: Study of immunological interaction between gamma delta T cells and the highly conserved protein in mycobacteria, HSP, heat shock protein will contribute to the elucidation of the mechanism of immunological surveillance and the mechanism of autoimmune diseases. In addition, it will also contribute to the development of a new mycobacterial vaccine which will give direct, protective immunity against tuberculosis.     

Correlation of tuberculin skin reaction with lymphocyte proliferation, interferon-gamma production and tumor necrosis factor-alpha production after in vitro stimulation with PPD and killed Mycobacterium tuberculosis using peripheral blood of healthy donors]

The quantitative relationships among the in vitro lymphocyte proliferation in peripheral blood in 19 healthy donors to purified protein derivative (PPD) and the killed Mycobacterium tuberculosis, interferon-gamma (IFN gamma) and tumor necrosis factor-alpha (TNF alpha) production in these culture supernatants, and the in vivo skin reaction to PPD which were simultaneously measured were studied. Statistical analysis was performed with t-test and multiple regression analysis: The results obtained were as follows; 1) The magnitude of the in vitro lymphocyte proliferation by PPD and the killed M. tuberculosis failed to correlate with the erythema and the induration of the in vivo skin reaction to PPD. 2) The erythema of skin test correlates with TNF alpha production in the culture supernatants that the lymphocytes in peripheral blood were cocultured with these antigens for 7 days. (R = 0.566062, 0.01 less than p less than 0.02) 3) There is a correlation between the erythema and the induration of skin test. (R = 0.526662, 0.02 less than p less than 0.05). 4) Though the magnitude of the lymphocyte proliferation to PPD correlates IFN gamma production in the culture supernatants (R = 0.525915, 0.02 less than p less than 0.05), these response to the killed M. tuberculosis correlates both IFN gamma production (R = 0.55049, 0.01 less than p less than 0.02) and TNF alpha production (R = 0.51283, 0.02 less than p less than 0.05) in the culture supernatants.(ABSTRACT TRUNCATED AT 250 WORDS)     

Influence of smoking on cutaneous delayed-type hyper sensitivity reaction by tuberculin skin test

Tuberculin test (purified protein derivative) is currently accepted as a standard investigation used in the diagnosis of tuberculosis (TB). Although the sensitivity of the test is reliable, a substantial number of those subjected to screening for TB by such test are cigarette smokers. This study is designed to investigate the effect of smoking on cell-mediated delayed-type cellular hypersensitivity (DTH) reaction by PPD. Prospective, case-control study was conducted at the Chest and TB unit of Chest Hospital Kuwait. The study population consisted of 357 healthy volunteers serving as controls and 200 in-patients under direct medical supervision and treatment for tuberculosis as cases. The mean age was 33.69 +/- 8.6 SD; 286 were current smokers and 271 were lifetime non-smokers. PPD test was done using 2TU RT23 SSI-Denmark on all subjects. Median PPD was significant among the cases (P=0.03) between smokers and non-smokers and was highly significant among the healthy controls (P<0.001). No significant difference was seen between median pack years of smoking and PPD levels among the patient group (P=0.264) but the difference was significant among the control group (P<0.001). Univariate analysis of variance (ANOVA) on PPD, taking into account age, pack years of smoking, ethnic groups and BCG scar showed sufficient response but was not statistically significant to all these factors. Smoking habit does not appear to influence the cutaneous delayed type hypersensitivity reaction by tuberculin skin test.     

The detection of antibodies to Mycobacterium tuberculosis and its reactivity to three different purified protein derivative by ELISA]

The specific PPD IgG antibodies in serum of patients with pulmonary tuberculosis and its reactivity to PPD of Mycobacterium tuberculosis M. bovis and M. avium were detected by ELISA. The results showed that the antibody level and positive rate of patients with pulmonary tuberculosis significantly increased than those of normal subjects. Specific antibody level and its positive rate were higher in hospitalized patients than those treated in clinic and both were higher in sputum positive cases than in sputum negative one. There was cross-reactivity among three different PPD. Absorption black test indicated that M. tuberculosis PPD had good specificity. PPD should be further purified in order to make ELISA as a useful serodiagnostic tool.     

Evaluation of DPPD, a single recombinant Mycobacterium tuberculosis protein as an alternative antigen for the Mantoux test

Although the tuberculin test has aided in the diagnosis of tuberculosis for more than 85 years, its interpretation is difficult particularly because sensitization with non-tuberculous mycobacteria leads to false positive tests. Using the guinea pig model of tuberculosis, we have recently described a recombinant antigen (DPPD) that could circumvent this problem. The DPPD gene is unique to the M. tuberculosis complex organisms and is absent in the organisms representative of all other members of the Mycobacterium genus. Moreover, DPPD induced strong DTH in 100% of the guinea pigs infected with M. tuberculosis and in none of the guinea pigs immunized with nine different species of Mycobacterium. Here we present results of a clinical investigation using DPPD. Mantoux test using both PPD and DPPD was initially performed in 26 patients with confirmed pulmonary tuberculosis and in 25 healthy PPD negative individuals. The results indicated that both PPD and DPPD elicited DTH in 24 out of the 26 patients. No DTH was observed in any of the PPD negative individuals. In addition, a small clinical trial was performed in a population of 270 clinically healthy and randomly selected individuals. DPPD produced a bimodal histogram of skin reaction size and PPD produced a skewed histogram. Because the DPPD gene is not present in non-tuberculous bacilli, these results suggest that this molecule can be an additional tool for a more specific diagnosis of tuberculosis.     

Absence of relationships between tuberculin responses and development of adult asthma with rhinitis and atopy

OBJECTIVE: To investigate the relationship between tuberculin skin responses and the development of adult asthma, rhinitis, and atopy. METHODS: Two hundred fourteen patients with mild-to-moderate asthma accompanied with rhinitis and 220 normal volunteers underwent a medical history, chest radiography, allergen skin-prick testing (SPT), bovine Mycobacterium tuberculosis vaccine (BCG) scar identification, purified protein derivative (PPD) tuberculin skin testing, serum-total and serum-specific IgE measurements, and bronchial provocation (provocative dose of histamine causing a 20% fall in FEV(1) [PD(20)]). RESULTS: Thirty-one normal volunteers (14.1%) and 168 asthma-rhinitis subjects (78.5%) had one or more positive skin test results (p < 0.0001). Neither the presence of a BCG scar nor a history of BCG vaccination had a significant effect on atopy in either group. The rate of PPD positivity had no statistical difference between atopy and nonatopy in both groups. In multivariate logistic regression analysis, the odds ratio for tuberculin reactivity was not related to the level of serum-total IgE nor to the level of serum-specific IgE to Dermatophagoides pteronyssinus (DP) and Dermatophagoides farinae (DF), skin response to DP and DF, and PD(20). Overall, no significant correlations were found between tuberculin skin reactivity and log serum-total IgE or PD(20). CONCLUSION: There is no relationship between history of tuberculosis infection, tuberculin responses, and development of adult bronchial asthma, allergic rhinitis, and atopy. Our study suggests that the protection provided by intradermal BCG vaccination in infants to prevent atopic diseases may be limited in early childhood, when a substantial memory of cellular immune modulation still exists.     

Attenuated response to purified protein derivative in patients with rheumatoid arthritis: study in a population with a high prevalence of tuberculosis

BACKGROUND: The purified protein derivative (PPD) skin test is the only widely used method which detects latent tuberculosis infection (LTBI) and is dependent on a normal T cell function. In rheumatoid arthritis (RA) the T cell function is altered, which may result in an inability to develop an adequate PPD reaction. OBJECTIVES: To evaluate the response to PPD in patients with RA and to compare it with that of control subjects. METHODS: 112 patients with RA and 96 healthy controls were studied. PPD 5 U was applied using the Mantoux method, and skin reaction was measured at 72 hours. The reaction was considered negative for PPD <5 mm. RESULTS: There were no significant differences in age, sex, history of bacille Calmette-Guerin vaccination, or tuberculosis contact between the two groups. The median size of the PPD induration in the patients with RA was significantly less than that in the control group (4.5 v 11.5 mm, p<0.01). 79 (70.6%) patients with RA compared with 25 (26%) of the control group had a negative reaction to PPD (p<0.01), a response not influenced by disease activity or duration of disease in the patients with RA. CONCLUSION: A PPD skin test is not an appropriate test for recognising LTBI in patients with RA in our population.