Search

Background Several destructive and immunotherapeutic methods are used in treatment of plantar warts, but an effective method with no or reduced recurrence has not been found till now. Objectives To evaluate the efficacy and safety of intralesional (IL) vitamin D3 (Vit.D3 ) injection vs IL tuberculin purified protein derivative (PPD) injection in the treatment of plantar warts. Methods Sixty patients with plantar warts were randomized into 3 equal groups: group I treated using IL tuberculin PPD every 2 weeks, group II treated using IL Vit.D3 every 4 weeks, and group III treated with IL saline every 2 weeks till complete clearance or for a maximum of 3 sessions. The follow-up period was 6 months. Results There was a statistically significant improvement in therapeutic groups than control with more significant improvement in group II than I. Regarding number of sessions required for complete response, there was a positive significant correlation in both groups, but more significant in group I. There was a negative correlation between the number of lesions and the response to treatment in both groups. Group II showed significantly better response to treatment in male patients. Both modalities were well tolerated, with no remarkable side effects and no recurrence in cured patients of both groups. Conclusions Both IL PPD and Vit.D3 injection are safe and effective for treatment of plantar warts even recalcitrant or multiple, with no postprocedural downtime, better results, and patient satisfaction. IL Vit.D3 injection has a superior advantage than PPD.

Cutaneous warts are a common pediatric complaint with modest response to first-line treatments. Warts are a manifestation of human papillomavirus (HPV) infection and are cleared by cell-mediated immunity (CMI). Intralesional immunotherapy treatments have been studied as alternative therapies, particularly for recalcitrant or multiple warts, including Candida antigen, mumps antigen, the combined measles, mumps, and rubella (MMR) vaccine, tuberculin purified protein derivative (PPD), and bacille Calmette-Guerin (BCG) vaccine. These treatments are thought to increase HPV recognition by stimulating CMI. In this review, we evaluate and compare the efficacy and adverse effects of intralesional immunotherapy in the treatment of pediatric warts. Articles met inclusion criteria if they specifically evaluated the effects of intralesional immunotherapy (candida, MMR, tuberculin PPD, or BCG) as treatment for cutaneous warts in a pediatric population, and if they quantified treatment effect in a reproducible manner. Twenty-one studies met criteria. Many studies demonstrated complete clearance of injected common warts in pediatric patients with clearance rates ranging from 23.3% to 95.2%. Distant wart resolution was common. Intralesional immunotherapy is a promising treatment option for cutaneous warts in children.

Background: Many destructive and immunotherapeutic modalities have been used for the management of plantar warts; however, an optimal treatment with high efficacy and absent or low recurrence has not been explored to date. Tuberculin purified protein derivative (PPD) shown promising results for treatment of viral warts without ablation. Aim of Study: To evaluate the efficacy and safety of intralesional (IL) PPD in the treatment of plantar warts. Patients and Methods: Twenty patients with plantar warts were included in this study and divided into two equal groups; group I was treated using IL tuberculin PPD every 2 weeks, group II was treated with IL saline every 2 weeks, till complete clearance or for a maximum of 3 sessions. The patients were followed-up for a period of 6 months. Results: There was a statistically highly significant im-provement in the therapeutic group compared with control group. This modality was well tolerated, with no remarkable side effects and no recurrence in cured patients during follow up period. A significant positive correlation existed between the number of sessions and treatment response. There was a significant negative correlation between the number of lesions and response to treatment. Conclusion: Intralesional PPD is safe and effective treat-ment alternative for the treatment of plantar warts even if they are recalcitrant or multiple, with no post-procedural downtime and better results and patient satisfaction.

Background: The immune system plays an important part in the clearance of molluscum contagiosum (MC) and, therefore, there has lately been a trend in using immunotherapy in MC therapy. Tuberculin-purified protein derivatives (PPDs) and topical cantharidin have not earlier been compared with their effectiveness in therapyAim: The aim of this research was to determine the effectiveness and security of intralesional immunotherapy in the therapy of MC with tuberculin PPD versus topical cantharidinPatients and methods: Twenty patients with various MC lesions received topical cantharidin as control (group A) and 20 MC patients received intralesional tuberculin PPD following prior intradermal immunity tests (group B‏).Results: Complete clearance of lesions was detected in 90.0% of patients in the cantharidin group; the partial response was detected in10.0% of the patients. However, in the PPD group, 85% of the patients showed a complete response and 15% showed a partial response, with no significant difference in the clinical response between the two groups. Mild side effects were detected‏.Conclusion: The results suggest that intralesional PPD and topical cantharidin 0.7% are effective and safe treatment modalities, but benefits of intralesional PPD is being a simple, effective and safe treatment with tolerable pain and can be an alternative treatment for multiple resistant types‏.

The Irish Bovine Tuberculosis (bTB) eradication programme operates under national legislation and fulfills OIE and EU trade requirements. Tuberculin purified protein derivative (PPD), a preparation obtained from the heat-treated products of growth and lysis of Mycobacterium bovis or Mycobacterium avium (as appropriate), is critical to the diagnosis of tuberculosis (TB). Standardization of Tuberculin PPD potency, the relative activity in sensitized animals compared to a reference standard, is essential to underpin the reliability of certification for international trade and to ensure that disease eradication programmes are effective and efficient. A Bovine International Standard Tuberculin PPD (BIS) was established by the WHO in 1986 and is used to determine comparative potencies of Tuberculin PPDs. Ideally, Tuberculin PPD potency should be evaluated in the species in which the tuberculin will be used but due to practical difficulties in performing potency assays in cattle, for routine PPD production, they are usually assayed in guinea pigs. Low potency tuberculin PPD is less efficient and thus inferior for bTB diagnosis. Difficulties experienced in the Irish bTB eradication programme have included the supply of sub-standard potency, and thus inferior, bovine (M. bovis) Tuberculin PPD in the late 1970s. The purpose of this paper is to outline the critical role of Tuberculin PPD assays carried out on naturally infected tuberculous cattle, as required by the OIE and under EU legislation in the quality control for the Irish Bovine Eradication Programme. Such assays ensure that the Tuberculin PPD used meets the diagnostic sensitivity and specificity requirements to underpin a successful national eradication programme.

Introduction and aim: Genital wart infection is a distressing sexually transmitted infection that causes marked fear and stress to both males and females. Tuberculin Purified protein derivative (PPD) injection is an upcoming safe and effective modality for the treatment of genital warts. Interleukin 17 (IL-17) was found to be lower in wart patients than controls. This study aimed at measurement of serum level of IL-17 in genital warts patients before and after PPD injection. Patients and methods: The study was carried out as case-control interventional study on 63 patients (37 males and 26 females) with genital warts attending Dermatology and Andrology Clinics, faculty of medicine, Suez Canal University hospitals. 50 controls were enrolled in the study. All patients have injected 10 tuberculin units of PPD intradermally in the mother or largest warts every 2 weeks till 6 sessions. Serum level of IL-17 was measured by Sandwich ELISA technique to all patients before and after PPD injection and to all control subjects. Results: The mean age of patients was 30.8 years, the mean duration of warts was 4.73 months, the mean number of warts was 9.76 warts and 14.3% of patients experienced side effects to PPD. (58.7%) of patients were completely recovered from warts. Mean serum IL-17 was 81.1 ± 64.2 pg/mL versus 118.9 ± 93.8 pg/mL in genital wart patients versus controls respectively (P < 0.05) and 81.1 ± 64.2 pg/mL versus 107.1 ± 66.8 pg/mL before and after PPD injection respectively (P < 0.05). Conclusion: IL-17 serum level was significantly lower in genital wart patients than normal controls. IL-17 serum level had been increased significantly after PPD intralesional injection than pretreatment levels and significantly related to clinical response to PPD. IL-17 suggested having a role in the clinical efficacy of PPD in the treatment of genital warts.

Background: Viral warts caused by human papilloma virus are a group of oncogenic viruses that result in verrucous growths on the skin and mucosae. Intralesional immunotherapy has been tried in recent years with success with the use of a variety of antigens to non-specifically stimulate the cell mediated immune response. Similarly, we undertook a study to evaluate the efficacy and safety of tuberculin purified protein derivative as an immunotherapeutic modality.Methods: A total of 25 patients were included in the study. Each patient was injected with 2.5 TU of tuberculin PPD (0.04 ml) intralesionally into most of the warts at 2 weekly intervals for a maximum of six sessions. It is an interventional study with follow-up upto 6 months. Descriptive analysis was done with use of SPSS version 23 tool. Complete resolution of warts was considered as the clinical end point. Response was graded as excellent response (>80% resolution), good response (50-80%), fair response (20-50%) and no response (Results: Out of the 25 patients, 14 patients showed excellent response (56%), 4 patients showed good response (16%), 5 patients showed fair response (20%) and 2 patients did not show any response (8%). Over 22 patients (88%) showed features of pain, erythema and inflammation at the injection site, whereas 3 patients (12%) developed itching at the site. The ‘faces’ pain rating scale was used to measure pain tolerance. An overall response rate (fair/good/ excellent) was given by 23 out of the 25 patients (92%), which proved satisfactory.Conclusions: Intralesional immunotherapy with PPD effectively cures warts which are present locally and also at distant sites. It is safe, economic and efficacious.

Introduction: Viral warts are caused by human papillomavirus. Although various treatment modalities are available, many of them result in the partial clearance of warts or recurrence. Immunotherapy has been tried in the recent times with variable success rates. We undertook a study to evaluate the safety and efficacy of immunotherapy using tuberculin purified protein derivative (PPD) for the treatment of viral warts. Materials and Methods: A total of 45 patients were included in the study. Each patient was injected with 10 TU of tuberculin PPD (0.1 ml) intralesionally in the largest wart at 2 weekly interval. A total of maximum six treatment sessions were conducted. Resolution of viral warts was considered as the clinical end point of the study. The clinical assessment was done by photographic measurements at baseline, before each treatment session, and 3 weeks after the completion of treatment. Results: A total of 62.2% patients (28 out of 45) showed complete clearance at injected and distant warts, eight patients (17.8%) showed partial clearance, and nine patients (20%) showed no improvement. No significant side effects were observed except for localized hair loss around injected viral wart over the scalp. Conclusion: Tuberculin PPD immunotherapy was found to be a safe and effective treatment modality for the treatment of viral warts.

Background: Viral warts are caused by human papillomavirus. There are innumerable treatment modalities but they may result in partial clearance or recurrence. Some newer and effective treatments include intralesional immunotherapy. We undertook a study to evaluate the safety and efficacy of two such immunotherapy namely tuberculin purified protein derivative (PPD) versus intralesional vitamin D3 for the treatment of viral warts.Methods: A total of 80 patients were included in the study. Group 1 patient were injected with 10 TU of tuberculin PPD (0.1 ml) intralesional while in group 2 patients were injected (0.5 ml) vitamin D3. The clinical assessment was done by photographic measurements at baseline, before each treatment session, and after the completion of treatment.Results: In group 1 A total of 80% (32 patients) showed complete clearance and 20% (8 patients) showed partial response while in group 2 72.5% (29 patients) showed complete clearance while 27.5% (11 patients) showed partial response. No significant side effects were observed except for pain at the site of injection.Conclusions: Intralesional vitamin D and intralesional Tuberculin PPD immunotherapy shows significant results in treatment of viral warts.

The manufacturers of the only tuberculin available up to now in Germany for intradermal TB tests according to Mantoux, Chiron Vaccines Behring, in 2004 unexpectedly stopped the production of the tuberculin Behring GT (GT="gereinigtes Tuberkulin"-purified protein derivative tuberculin). Only residual stocks were sold during the preceding months. The stocks of GT 10 were already depleted at the beginning of 2005, while there are small supplies left of GT 100 and GT 1000. As a temporary solution, Chiron Vaccines Behring is offering to import the Italian tuberculin Biocine PPD 5 IE lyophil produced by Chiron S.r.l. in Siena. As this is not licensed for sale in Germany, it is necessary to obtain an exceptional prescription (Einzelverordnung) according to section sign 73 (3) of the Federal Law Relating to the Manufacture and Distribution of Medicine (Arzneimittelgesetz, AMG). In the long term, Chiron Vaccines Behring plan to secure the supply of tuberculin in Germany by importing, starting in the summer of 2006, the tuberculin produced by Chiron Vaccines Evans in the UK (PPD Evans). However, these plans involve changing over to a different type of tuberculin twice within a very short period of time. Another problem is the unresolved issue of bioequivalence. Besides the above-mentioned tuberculins produced by Chiron Vaccines, a further possibility would be the import of the tuberculin PPD RT23 SSI of the Statens Serums Institute (Copenhagen/Denmark), which is recommended by the World Health Organization (WHO) as the standard tuberculin and which has already been introduced in several European countries, or of other tuberculins such as Tubersol (PPD CT68), which is used in the US. Together with the Robert Koch Institute (RKI) and the Paul Ehrlich Institute (PEI), the German Central Committee against Tuberculosis (DZK) is striving to find a solution, in view of the urgent need for an uninterrupted supply of tuberculin in Germany for diagnostic purposes and contact tracing. A uniform tuberculin should be used in all German regions as a basis to secure a standardized testing procedure and national comparability of test results. The estimated annual requirement for Germany is two million tuberculin test doses. The Statens Serum Institute is currently evaluating the possibility of licensing PPD RT23 SSI in Germany. Chiron Vaccines Behring communicated that it is not aiming to have the Biocine tuberculin licensed for Germany but intends to apply for a license for the tuberculin PPD Evans. Sanofi Pasteur MSD at this point does not intend to have a tuberculin like Tubersol (PPD CT68) licensed for Germany. Until a licensed tuberculin is again available in Germany, it would be very useful if, especially in settings where stocks of tuberculin are essential (e.g. public health services or hospital dispensaries), an exceptional import license could be obtained.

Since 1951, the tuberculin PPD-S1 has been used to standardize commercial PPD reagents and perform special tuberculin surveys. PPD-S1 is now in short supply and a new standard (PPD-S2) has been manufactured. To determine if PPD-S2 is equivalent and can replace PPD-S1, we conducted a double-blind clinical trial. Between May 14 and October 28, 1997, 69 subjects with a history of culture-proven tuberculosis (TB patients) and 1,189 subjects with a very low risk for TB infection were enrolled, received four skin tests (with PPD-S1, PPD-S2, and one each of the commercially available PPDs), and had reactions measured by two trained observers. Among the TB patients, we found statistically indistinguishable immunogenicity (mean reaction size +/- standard deviation): 15.6 +/- 6.6 mm for PPD-S1 and 14.8 +/- 5.6 mm for PPD-S2. Among low-risk subjects, the tests had equally high specificities (PPD-S1, 98.7% and PPD-S2, 98. 5%), using a 10-mm cutoff. The number of discordant (negative versus positive) interpretations for PPD-S2, assuming that low-risk subjects who had a/= 10 mm reaction to PPD-S1 were truly infected, was low (0.5%) and indistinguishable from the rate of discordant interpretations of the same test when read by two different observers (0.8%). The study results indicate that PPD-S2 is qualified to be used as the new U.S. reference standard for PPD tuberculin.

Ireland currently obtains its avian and bovine tuberculin purified protein derivatives (PPDs) from a single source. Because problems of supply or quality cannot be discounted, it is prudent that Ireland identify alternative supplier(s) as part of a broad risk management strategy. Therefore, the aim of this study was to compare the performance of a number of different tuberculin combinations (that is, pairings of bovine and avian PPD; with different manufacturers) in the single intradermal comparative tuberculin test (SICTT), as currently performed in Ireland. The study was randomised, controlled and double-blinded. A total of 2172 cattle were used in the study. Each animal was tested using two SICTTs, the first based on the tuberculin combination in current use, and the second using one of six trial tuberculin combinations. Analyses were conducted to compare both reactor-status and skin increase. For each control/trial tuberculin combination, there was good agreement between the control and trial reactor-status. Differences in skin increases were mainly confined to animals categorised as either negative or severe inconclusive. However, the measured differences were minor, and unlikely to have a significant impact on the actual test outcome, either for individual animals or for herds. In conclusion, while further studies determining sensitivity and specificity in Ireland would have to be done in the event of a change in tuberculin PPD there should be minimal disruption of the national programme if alternative tuberculin PPDs meeting WHO, OIE and EU regulations were used. In this study, the precision of the guinea pig bio-assay to assess tuberculin potency was low and therefore Ireland should maintain its practice of periodically assessing potency in naturally infected cattle, even though this is not currently required under WHO, OIE or EU Regulations.

In national bovine tuberculosis (BTB) control programmes, testing is generally conducted using a single source of bovine purified protein derivative (PPD) tuberculin. Alternative tuberculin sources should be identified as part of a broad risk management strategy as problems of supply or quality cannot be discounted. This study was conducted to compare the impact of different potencies of a single bovine PPD tuberculin on the field performance of the single intradermal comparative tuberculin test (SICTT) and single intradermal test (SIT). Three trial potencies of bovine PPD tuberculin, as assayed in naturally infected bovines, namely, low (1192 IU/dose), normal (6184 IU/dose) and high (12,554 IU/dose) were used. Three SICTTs (using) were conducted on 2102 animals. Test results were compared based on reactor-status and changes in skin-thickness at the bovine tuberculin injection site. There was a significant difference in the number of reactors detected using the high and low potency tuberculins. In the SICTT, high and low potency tuberculin detected 40% more and 50% fewer reactors, respectively, than normal potency tuberculin. Furthermore, use of the low potency tuberculin in the SICTT failed to detect 20% of 35 animals with visible lesions, and in the SIT 11% of the visible lesion animals would have been classified as negative. Tuberculin potency is critical to the performance of both the SICTT and SIT. Tuberculin of different potencies will affect reactor disclosure rates, confounding between-year or between-country comparisons. Independent checks of tuberculin potency are an important aspect of quality control in national BTB control programmes.

Non-BCG-vaccinated schoolchildren (8 or 9 years of age) were simultaneously tested on separate arms with 2 IU PPD RT23 and 0.1 microgram Mycobacterium avium sensitin RS10 or 0.1 microgram Mycobacterium scrofulaceum sensitin RS95. None of the 2819 analysed children had any known exposure to tuberculosis. A total of 3.4% reacted with an induration greater than or equal to 6 mm to PPD RT23. Half the number of children were tested with M. avium sensitin and 25.4% reacted while the remaining were tested with M. scrofulaceum sensitin and 32.4% reacted when the cut-off was 6 mm. For about 90% of the children the sensitin reaction was larger than or equal to the tuberculin (PPD RT23) reaction. Correlation analyses showed that moderate and high PPD RT23 values were combined with still higher sensitin values, indicating that the tuberculin reactions were mainly cross-reactions due to the antigenic similarity between tuberculin and sensitins. The presence of birds, dogs and cats in the homes was combined with an increased frequency of children reacting to the sensitins used. The children with reactions to PPD RT23 greater than or equal to 6 mm were examined and chest X-rays were performed. None of them showed any signs or symptoms of mycobacterial disease. In non-BCG-vaccinated Swedish schoolchildren without clinical signs of tuberculosis and without known contact with a contagious tuberculous person, indurations less than 12 to 14 mm on tuberculin testing are probably caused by atypical mycobacteria. In such cases sensitin tests should be performed to verify the suspicion.

The aim of this study was to determine the epidemiological link between tuberculosis in livestock and man in a livestock rearing community. This was done through a follow up of human patients, diagnosed with clinical tuberculosis at the Sokoto Specialist Hospital, to their residences and carrying out tuberculin test on their domesticated animals and human contacts. From the human patients, tubercle bacilli were isolated through bacteriological examination while the single cervical comparative tuberculin test was conducted for the animals. Findings revealed that mycobacterium species isolated from the human patients were M. tuberculosis 9; M. bovis 4 and atypical mycobacterium 3. Patients' relation reacted to Mantoux test showing 50% infection rate. The comparative tuberculin test conducted showed that 4 reacted to M. tuberculosis 5 to M. bovis and 7 no reaction. The animals kept in the household of the patients showed reaction to tuberculin PPD which suggest that the causative organism in the patients and that infecting the animals are the same species. There is a need to undertake an in-depth study into transmissibility of tuberculosis between livestock and man within a household. Keywords : In-contact, Mycobacterium Infection, Cattle, Tuberculosis Patients Nigerian Veterinary Journal Vol. 27 (3) 2006 pp. 87-90

We have evaluated the comparative intradermal skin (CID) test, the interferon gamma (IFN- γ ) assay, two ELISAs with bovine PPD antigen, the standard and the anamnestic using sera obtained, respectively, at the time of the tuberculin injection and 15 days later, for the diagnosis of caprine tuberculosis (TB). The sensitivity and specificity results were high for the CID test (83.7%, 100%), the IFN- γ assay (83.7%, 96%) and the anamnestic ELISA (88.6%, 95.8%). In contrast, they were comparatively low for the standard ELISA (54.9%, 88%). However, test results with the standard ELISA were positive in a group of goats with cavitating TB (100%). A combination of the CID test and the IFN- γ assay offered the highest sensitivity, 95.8%, and also high specificity, 96%. In spite of this, the evidence that the serological tests were most sensitive for the detection of goats with severe lesions (100% positivity) suggested that a combination of CID test and anamnestic ELISA may be most useful as part of an eradication campaign against caprine TB.

Background : Tuberculin skin test is a method to examine M. tuberculosis infection and has been used all over the world. But various factors make it difficult to understand testing results. In 2000, the American Thoracic Society recommended that skin test results should be decided by considering risk factors of the tested. In Korea, high tuberculosis infection rate and BCG vaccination rate make it difficult to differentiate current infection, past infection, and no infection by the skin test. This study was attempted to examine a negative predictive value of the skin test to understand how the skin test acts on deciding administration of anti-tuberculosis drug. Methods : From Mar. 1 to Jul. 31 in 2001, the test was performed for patients hospitalized in Department of Internal Medicine, Hallym University College of Medicine, Chunchon, Korea by administering Tuberculin PPD RT23 2 TU (0.1 ml)to them that has been currently used in Korea based on Mantoux method. They were decided to be infected with tuberculosis bacilli by following diagnostic standard: 1) tuberculosis bacilli was cultured in sputum by microbiological diagnostic standard or Acid-fast bacilli was proven on a microscopic examination or 2) tuberculosis bacilli was not proven in the aforesaid microbiological test by clinical diagnostic standard, while there was opinion or symptom suitable for tuberculosis by radiographic or histological standard so the doctor decided to apply the tuberculosis treatment. Results : In this study, total 210 patients except 20 patients (8.7%) among 230 hospitalized patients were evaluated. Their average age was 60±16.8 years, and male-female rate was 1.28 : 1 (male: 118, female: 92). Number of patient, who was diagnosed and decided as tuberculosis, was 53(25.2%). Pulmonary tuberculosis was found in 45 patients (84.9%); 22 patients were decided to be positive in the Acid-fast bacilli smear test by microbiological examination (culture positive: 13, culture negative: 9), and 23 patients were decided to be tuberculosis patients by clinical diagnosis standard. Tuberculosis pleuritis was found in 8 patients (15.1%); 4 patients were diagnosed and decided by histological standard, and 4 patients were decided and treated by clinical standard. In differentiating patients into 'Negative' and 'Positive' by the skin test standard of the American Thoracic Society, negative predictive value 92.3%, positive predictive value 47.3%, sensitivity and specificity were 83%, 68.8%, respectively. Conclusion : In hospitalized respiratory patients, there was high negative predictive vlaue 92.3% by tuberculin skin test, therefore skin test would be a important factor for deciding administration of anti-tuberculosis drug on negative skin test patient.

The Pan American Health Organization, through its Pan American Zoonoses Center (CEPANZO), distributes the concentrated stock solution of PPD batch RT23 (1 mg ml−1 = 50 000 TU ml−1) to national central laboratories. In the laboratories the stock solution is diluted to 2 TU/0·1 ml and dispensed with the addition of Tween 80 and either chinosol or phenol as preservatives in rubber-stoppered vials for use in the Mantoux test. The expiry date is usually fixed at six months after filling. Samples of nine lots of PPD 2 TU from seven countries (Argentina, Brazil, Colombia, Dominican Republic, Ecuador, Paraguay and Uruguay) were received by CEPANZO for quality control. The biological activity and phenol concentrations of these samples were assayed periodically after storage at 4, 25 or 37 °C either unopened or opened and re-used in a manner simulating extreme field conditions of use and storage. The object of the study was to determine the stability of the biological activity as well as the maintenance of sterility in these conditions with a view to assessing the need for stricter requirements for the labelling of tuberculin. The results obtained showed that PPD 2 TU with 0·4–0·5% (wv) phenol added, and dispensed in vials, was stable in its biological activity and maintained its sterility. Therefore, it seems unnecessary to issue stricter requirements than those currently applied. PPD 2 TU can be used for six months after dilution provided that it has been stored at 5 ± 3 °C, protected from sunlight, and held in vials or ampoules filled to capacity provided that containers used twice with a remaining volume of half or less of the original volume were discarded.

A reference system for the soluble antigens of Mycobacterium bovis, strain BCG is described. The system is based on crossed immunoelectrophoresis with intermediate gel. A commercially available immunoglobulin preparation made from rabbit anti-BCG hyperimmune serum was used as reference antibodies, while a concentrated BCG culture filtrate was used as reference antigen. The pattern obtained was highly reproducible, and most of the components were stable when the fiftyfold-concentrated culture filtrate was stored at -20 degrees C. About thirty different antigenic components were selected as reference antigens and numbered. The majority of the reference antigens were present in extracts prepared from BCG by ultrasonication or bacterial press extraction. Use of the system for studies of antigenic relationship between mycobacteria, identification and quantification of antigens, and characterization of antimycobacterial antibodies are illustrated by examples. The antigens of two preparations of tuberculin purified protein derivative (PPD) were identified. The antigen designated BCG60 was found to be a najor constituent of tuberculin PPD. Evidence is presented that this antigen is cell wall associated.

The biological potency of 15 commercially available tuberculin solutions prepared from a master batch of tuberculin purified protein derivative (PPD) (PPD-CT68) and using a newly formulated diluent (Connaught diluent) containing 0.0005% Tween 80 as an anti-adsorption agent and 0.3% phenol as a preservative, was determined after storage for various intervals at 4, 24 and 37 degrees C. The 5 tuberculin units (TU) per 0.1 ml dose solutions were bioequivalent to a non-stabilized solution of PPD-S whereas the 1 TU and 250 TU per dose solutions were equivalent by calculation to a non-stabilized solution of PPD-S. It was found that the PPD solutions of all 3 strengths, 1, 5 and 250 TU per dose, were stable for at least 3 years at 4 degrees C and for 2 years at room temperature (24 degrees C). Even at 37 degrees C the solutions of all 3 strengths were stable for at least 1 year. The stability of Connaught tuberculin PPD solutions has not been affected by the changes in formulation. The stability data suggest that the expiry date of the newly formulated tuberculin products could be at least two years from the data of the last satisfactory potency test. Although these products are stable for at least 1 year even at 37 degrees C, we nonetheless agree with the Canadian and U.S. regulations that they be stored at 2 to 8 degrees C in their original containers.

The biological activity of purified protein derivative (PPD) of mammalian tuberculin was compared with that of old tuberculin (OT) prepared from the same strains of Mycobacterium tuberculosis . The tests were carried out intradermally in groups of experimentally sensitized guinea-pigs and in naturally sensitized cattle. Tuberculin PPD was found to be more specific than OT. Comparisons against tuberculin PPD of the precipitable and non-precipitable nitrogen fractions of OT showed that there are qualitative differences in the proteins of PPD and OT, apparently the result of the prolonged heating of OT during processing. The proteins of OT were less potent per unit of weight and were also less specific than those of PPD. Concentrated solutions of tuberculin PPD and of OT stored in full containers at 4° C. in the dark retained full potency after 10 years. Adsorption of active material to the walls of glass containers from tuberculins in high dilution resulted in greater losses in potency of OT than of tuberculin PPD. It is concluded that there is no reason for the continued use, under any conditions, of OT instead of tuberculin PPD

A technique has been described whereby tuberculin PPD is coupled, via tetrazotized benzidne, onto the surfaces of formalinized red blood cells. A stable antigen preparation is thereby obtained, which may be stored at –40°C. The purpose of the present method is to facilitate the study of antibody to tuberculoprotein.

This article deals with the interaction bet ween the preservative 8-hydroxyquinoline sulfate (8-HQS) and the surface-active agent polysorbate 80. In buffered solution (pH 7.38) 8-HQS is dissociated to 8-hydroxyquinoline (8-HQ) and H 2 SO 4 , and it is the base 8-HQ that forms a reversible association with polysorbate 80. The degree of binding of 8-HQ to polysorbate 80 was shown to be a function of the concentration of the nonionic surface-active agent. Polysorbate 80 at low concentration (about 0.00 5 per cent) has practically no effect on the concentration of 8-HQ in a buffered solution (pH 7.38) as used for preparing dilutions of tuberculin PPD for the intracutaneous met hod (Mantoux test). 8-HQS, in a buffered solution of pH 3, does not interact with polysorbate 80.

A retrospective study of a group of persons skin-tested during the period, 1947 to 1961, in the metropolitan Kansas City area has shown that probability of breakdown with clinical disease increases with size of reaction to intermediate strength tuberculin PPD-S. However, the number of cases observed in the group was lower than that generally experienced in the area. Factors which might explain this include the following: (1) age composition of the group, which was for the most part made up of school children; (2) loss from study group by emigration from study areas; and (3) possibility that cross reactions were induced by infection with atypical mycobacteria, although this is an area of rather low prevalence of these infections in children.

A serological technique, using formalinized red cells coupled to tuberculin PPD as antigen, has been assayed in a trial series of 124 tuberculous and 139 non-tuberculous patients. The stability of the antigen preparation, and the omission of an adsorption procedure for the removal of isoagglutinins facilitated the performance of the test. The results indicate that the technique warrants further trial as a diagnostic criterion of infection tuberculosis.

We evaluated delayed hypersensitivity in humans to three soluble an < tigens (tuberculoprotein, coccidioidin, and Candida extract) with the leukocyte migration inhibition test. Skin reactivity and migration inhibition correlated to a high degree with all the antigens. Cell migration was not inhibited in subjects without a positive skin test or clinical history. Sensitivity to tuberculin measured in vitro after vaccination with Bacillus of Calmette and Guerin correlated well with 25 tuberculin PPD skin test units, and indicates that the LMI test is more sensitive in detecting cellular hypersensitivity than skin tests with intermediate strength tuberculin PPD.

Two studies were performed to compare the disposable liquid tuberculin PPD 'Imotest' with the Mantoux test. While there was a tester effect in the first study indicating the need for accurate application of the imotest, the results of the second study showed that the two tests were comparable in selecting truly positive and negative tuberculin reactors. Results were compared both by conventional methods and by a method new to tuberculin testing, the maximum likelihood technique. This method gives a more accurate estimate of false reaction rates. A significant difference was found between reading at 48 and 72 h for both tests. The acceptability and ease of reading of the imotest, with a tuberculin selectivity comparable to the Mantoux method, make it a suitable alternative tuberculin test, especially for screening large populations.

Lactoferrin and avidin are progesterone-inducible glycoproteins in the human and chicken reproductive tracts, respectively. The effect of these proteins on lymphocyte proliferation was studied. The results showed that both proteins suppressed Concanavalin A (Con A)-induced responses so that a 10(3)-fold higher concentration of Con A was required for an optimal response. In contrast, avidin and lactoferrin had no effect on lymphocyte proliferation induced by other mitogens, tuberculin PPD or allogeneic cells. The effect of lactoferrin and avidin on lymphocyte proliferation seem to be caused by their binding to Con A. The significance of these findings is discussed.

The stability of a dilute solution of tuberculin purified protein derivative (PPD) at extreme temperatures has been studied. It was found that tuberculin PPD retained its potency for a period of at least 91 days at −28°C. However, the same preparation stored at 60°C showed some evidence of a loss of potency as early as 5 days after storage, and its overall potency declined up to 47 days, and then remained constant. No loss of phenol was evinced at −28°C, whereas, at 60°C there was a significant loss of phenol.

We have demonstrated progressive glomerular sclerosis during the years following subsidence of the acute manifestations of poststreptococcal glomerulonephritis (PSGN). Since evidence of a humoral-immune basis for this progression is lacking, we have examined cell-mediated immunity directed against streptococcal protoplast membrane antigens as a possible pathogenetic mechanism. Lymphocyte transformation, utilizing highly purified streptococcal antigens from types 12 and 57 (nephritogenic strains) and type 11 (rheumatic strain), was measured by [ 14 C] thymidine incorporation in 12 patients with evidence of persistent disease 4 to 18 years after acute PSGN, and in 12 normal controls. Dose response curves with nephritogenic strains, as well as with the rheumatic strain, showed a depressed response in PSGN, especially at higher concentrations of antigen. There was no difference between the two groups in blastogenic response to phytohemagglutinin-M (PHA-M), tuberculin PPD, and streptokinase-streptodornase (SKSD). The paradoxically depressed cellular-immune response to streptococcal protoplast membrane antigens in persistent PSGN may be causally related to development of the disease rather than to the occurrence of chronicity.

Summary Depot tuberculin (PPD) creams containing 5 and 2 mg. PPS/g. or ml. have been used for multiple puncture tests in man and were investigated: ( a ) for their ability to produce skin test reactions in comparison with Mantoux tests, and ( b ) for their capacity to persist locally in the skin for prolonged periods. The depot PPD creams produced positive reactions in all Mantoux positive subjects and in many of those who did not react to Mantoux tests with 100 tuberculin units. Out of a total of 521 young adult and adult subjects, 92 per cent gave positive reactions to the most satisfactory depot PPD ‘Eucerin’ cream (5 mg./g.), compared with 80 per cent of positive reactions to the Mantoux tests. No reactions were produced by depot PPD ‘Eucerin’ cream tests in 49 infants and children up to 14 years of age. Intracutaneous injections of live BCG vaccine were given to 15 subjects who reacted only to the depot PPD cream test, and 14 of those gave accelerated reactions, thus confirming the immunological significance of the low degree of sensitivity demonstrated by the depot PPD cream test. Mantoux tests performed after the BCG vaccination in these subjects showed that their tuberculin sensitivity had increased. Positive reactions to tests with the depot PPD cream were produced in Mantoux negative patients with sarcoidosis, and have been reported in tuberculin desensitized patients who did not react to Mantoux tests with 10,000 tuberculin units of Old Tuberculin. Local persistence of the depot PPD cream was shown by the appearance, after BCG vaccination, of positive reactions at the site of previously negative tests, performed three weeks before the vaccination, thus demonstrating tuberculin conversion from negative to positive. The positive depot PPD cream test reactions were always of mild intensity and persisted for at least as long as twelve weeks. The value of a potent, locally persistent depot PPD cream for multiple puncture test is discussed, with particular reference to the significance of low degrees of tuberculin sensitivity.

SummaryDiscrete pools of tuberculopoly-saccharide and tuberculoprotein antigens, prepared by continuous-flow electrophoretic separation of an unheated culture filtrate of M. tuberculosis (H37Rv strain), were tested for induction of lymphocyte blastogenesis in cultures of blood leukocytes from healthy individuals who reacted to an intermediate skin test dose of tuberculin PPD. Approximately 80% of the blastogenic activity of the unseparated culture filtrate was found in the pool of protein antigens while only 7% was detected in the polysaccharide pool. As indicated by immunoelectrophoretic and chemical analyses, the minimal activity of the pooled polysaccharides was attributed to their trace contamination by the protein antigens. It was concluded that tuberculin-induced lymphocyte blastogenesis reflects cellular reactivity to tuberculoprotein antigens.The competent technical assistance of Frances E. Soehnlen is gratefully acknowledged.

The presence of 8-hydroxyquinoline sulfate (8-HQS) as a preservative in tuberculin purified protein derivative (PPD) solutions leads to the formation of a dark crystalline deposit. In this investigation it has been shown that the deposit was due to chelates resulting from the reaction of metals with 8-hydroxyquinoline (8-HQ) liberated from its sulfate. The metal impurities in tuberculin solution originate mainly from the buffer salts and the PPD. The removal of these metal impurities from the tuberculin PPD did not affect the potency of the preparation. Previous workers have shown the need to maintain the metal content within fairly narrow limits in order to render 8-HQ effective as a preservative. The necessity to keep the metal content within narrow limits in order to render the preservative effective, and to avoid the development of organo-metallic deposits, was considered an important drawback to its usefulness as a preservative.

The method used to produce “Connaught” tuberculin purified protein derivative (PPD) is described. The tuberculin PPD for the multiple-puncture method was shown to be stable for at least 24 months at 5 C; tuberculin PPD for the intracutaneous method was shown to be stable at 5 C and 24 C for a period of 18 months in the presence of Tween 80. Evans blue or brillant vital red was added to tuberculin PPD for improved testing by the multiple-puncture method. These tinted tuberculin preparations were found to be as stable as the Connaught tuberculin PPD preparations without dye at 5 C. Freeze-dried tuberculin PPD with Plasdone as an inert base was found to be remarkably stable for a period of at least 24 months at 5, 24, and 37 C.

Two hundred and thirty-seven chronically sick aged patients were skin tested with Tuberculin PPD, Intermediate Test Strength. Healthy younger staff members constituted a control group. The percentage of negative reactors among the patients appeared to be directly related to the severity of their chronic illness, as represented by the various ward categories. The percentage of negative reactors among the patients who subsequently showed a positive reaction to Tuberculin PPD, Second Test Strength, was directly related to the state of health of the patients. In sick old persons, a negative reaction to Tuberculin PPD, Intermediate Test Strength, should be followed by testing with Second Test Strength before it can be confirmed that the person is a negative reactor. The significance of a negative reaction to even very large doses of Tuberculin PPD should be viewed in the context of the overall clinical picture when dealing with a chronically sick aged patient.

Introduction: Guidelines recommend storing tuberculin purified protein derivative (PPD) refrigerated. However, especially in developing countries, maintaining the product refrigerated under field conditions can be difficult, limiting its use. Here we determine the effect of prolonged exposure to high temperatures on the potency of human, bovine and avian tuberculin PPD. Methodology: Human, bovine and avian tuberculin PPD were stored for several weeks exposed to temperatures ranging from 37º to 100ºC. The potency was evaluated in vivo, in sensitized or naturally infected animals. Results: Most test situations didn't affect the biological activity of the tuberculin PPDs and only very long and extreme incubations (several days at 100°C) compromised the potency. Conclusions: Tuberculin PPD is very stable and can be stored or transported for long periods without refrigeration.