Purified Protein Derivative Of Tuberculin Research Articles

Efficacy and Safety of Intralesional Injection of Tuberculin PPD in Treatment of Plantar Warts

Abstract 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.

Targeting of tolerogenic dendritic cells to heat-shock proteins in inflammatory arthritis

BackgroundAutologous tolerogenic dendritic cells (tolDC) are a promising therapeutic strategy for inflammatory arthritis (IA) as they can regulate autoantigen-specific T cell responses. Here, we investigated two outstanding priorities for clinical development: (i) the suitability of using heat-shock proteins (HSP), abundant in inflamed synovia, as surrogate autoantigens to be presented by tolDC and (ii) identification of functional biomarkers that confirm tolDC regulatory activity.MethodsCell proliferation dye-labelled human peripheral blood mononuclear cells of IA (rheumatoid arthritis (RA) and psoriatic arthritis (PsA)) patients or healthy donors were cultured with HSP40-, HSP60- and HSP70-derived peptides or recall antigens (e.g. tuberculin purified protein derivative (PPD)) in the presence or absence of tolDC or control DC for 9 days. Functional characteristics of proliferated antigen-specific T-cells were measured using flow cytometry, gene expression profiling and cytokine secretion immunoassays. Repeated measures analysis of variance (ANOVA) with Bonferroni correction for comparisons between multiple groups and paired Student t test for comparisons between two groups were used to determine significance.ResultsAll groups showed robust CD4+ T-cell responses towards one or more HSP-derived peptide(s) as assessed by a stimulation index > 2 (healthy donors: 78%, RA: 73%, PsA: 90%) and production of the cytokines IFNγ, IL-17A and GM-CSF. Addition of tolDC but not control DC induced a type 1 regulatory (Tr1) phenotype in the antigen-specific CD4+ T-cell population, as identified by high expression of LAG3, CD49b and secretion of IL-10. Furthermore, tolDC inhibited bystander natural killer (NK) cell activation in a TGFβ dependent manner.ConclusionsHSP-specific CD4+ T-cells are detectable in the majority of RA and PsA patients and can be converted into Tr1 cells by tolDC. HSP-loaded tolDC may therefore be suitable for directing T regulatory responses to antigens in inflamed synovia of IA patients. Tr1 markers LAG3, CD49b and IL-10 are suitable biomarkers for future tolDC clinical trials.

Detection of native interferon-γ in nyala (Tragelaphus angasii): Towards diagnosing tuberculosis.

Mycobacterium bovis is the main cause of tuberculosis in wildlife. In South Africa, African buffaloes (Syncerus caffer) are a wildlife maintenance host while a number of other species are considered spillover hosts. Nyala (Tragelaphus angasii), a large antelope species from Southern Africa, is frequently traded and can be infected with M. bovis. Interferon gamma (IFN-γ) release assays that detect cell-mediated immune (CMI) responses to M. bovis infection have shown promise in elephants, rhinoceroses and buffaloes. The BOVIGAM® assay is a commercial IFN-γ release assay designed to detect tuberculosis in cattle and has been validated in buffaloes. We tested the suitability of the BOVIGAM® assay to detect native IFN-γ release in nyala. Blood samples collected from 17 nyalas were stimulated with different mitogens and IFN-γ release measured. We found that incubating whole blood with phorbol 12-myristate 13-acetate and calcium ionophore (PMA/CaI) resulted in the highest levels of IFN-y release. Samples stimulated with tuberculin purified protein derivatives of M. bovis (PPDb) and M. avium (PPDa) did not show significant IFN-γ production. An intradermal tuberculin test (IDT) and culture of tissues from 15 of the 17 culled nyala were also performed, which supported the findings of the BOVIGAM® assay, suggesting the potential value of this assay for the diagnosis of tuberculosis in nyala.

Differed IL-1 Beta Response between Active TB and LTBI Cases by Ex Vivo Stimulation of Human Monocyte-Derived Macrophage with TB-Specific Antigen.

Background The difference of macrophage-specific interleukin-1 beta (IL-1b) response between latent tuberculosis infection (LTBI) and active tuberculosis (TB) remains less studied. Method We performed this prospective study and recruited active TB patients, contacts with LTBI, and uninfected contacts. The gene and protein expression of human monocyte-derived macrophage (hMDM) after ex vivo stimulation by early secretory antigenic target-6KD (ESAT-6) and tuberculin purified protein derivatives (PPD) was studied by real-time PCR and flow cytometry. The effect of caspase-1 inhibitor was also studied. Result The IL-1b gene expression after 6 hr ESAT-6 1 μg/ml stimulation was different among active TB patients (n = 12), LTBI cases (n = 12), and uninfected contacts (n = 23) (log fold change: 0.98 ± 1.26 vs. 2.20 ± 0.96 vs. 2.20 ± 0.96, P = 0.013). The IL-1b gene expression at 24 hours was higher than that at 6 hours in LTBI cases (n = 4) and uninfected contacts (n = 6). After 24 hr ESAT-6 1 μg/ml stimulation, the percentage of IL-1b-expressed hMDM was borderline lower in the active TB patients (n = 9) than in the LTBI cases (n = 10) (14.0 ± 11.2% vs. 31.6 ± 22.5%, P = 0.065). Compared with ESAT-6 1 μg/ml stimulation but without the addition of caspase-1 inhibitor (CasI) (55.6 ± 16.3%), the percentage of IL-1b-positive hMDMs decreased after addition of CasI (50 μg/ml CasI: 49.8 ± 18.2%, P = 0.078; 100 μg/ml CasI: 46.6 ± 20.8%, P = 0.030; 150 μg/ml CasI: 33.7 ± 15.5%, P = 0.016). Conclusions This study revealed that macrophage-specific IL-1b response differed among different stages of Mycobacterium tuberculosis infection. The role of IL-1b and inflammasome in the process of LTBI progressing to active TB warrants further investigation.

Tuberculin PPD Potency Assays in Naturally Infected Tuberculous Cattle as a Quality Control Measure in the Irish Bovine Tuberculosis Eradication Programme.

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.

CDC releases recommendations amid shortage of tuberculin skin test antigens

In June, CDC announced a nationwide shortage of Aplisol (Par Pharmaceuticals), a purified-protein derivative (PPD) tuberculin antigen. According to the agency, the shortage is expected to last 3 to 10 months, with a supply interruption of Aplisol 5 mL (50 tests) beginning in June 2019 and Aplisol 1 mL (10 tests) in November 2019 or sooner if demand increases before November. Aplisol is one of two PPD tuberculin antigens approved by FDA to perform tuberculin skin tests (TST) for the detection of latent Mycobacterium tuberculosis (TB) infection. The other is Tubersol (Sanofi Pasteur). Amid this shortage, CDC has released the following recommendations to mitigate the limited supply of Aplisol and prevent a decline in TB testing capability:▪Substitute interferon-gamma release assay (IGRA) blood tests for TSTs.▪Substitute Tubersol for Aplisol.▪Prioritize allocation of TSTs to at-risk groups in consultation with state and local public health authorities. According to the agency, the high-risk groups that take priority are as follows:▪People who have been recently exposed to persons with TB disease▪People born in or who frequently travel to countries where TB disease is common▪People who currently live or used to live in large group settings such as homeless shelters or correctional facilities▪People with weaker immune systems, such as those with certain health conditions or taking medications that may alter immunity▪Children, especially those younger than age 5 years, if they are in one of the risk groups mentioned above With the release of these new recommendations, CDC assured providers that the overall concordance among the different tests for TB disease is high. However, the agency cautioned that “switching between PPD skin test products or between TSTs and blood tests in serial testing may cause apparent conversions of results from negative to positive or reversions from positive to negative.” The agency explained that this may be due to “inherent inter-product or inter-method discordance, rather than change in M. tuberculosis infection status.” False-positive reactions may occur with TSTs. Causes may include, but are not limited to, infection with nontuberculosis mycobacteria, previous bacilli Calmette-Guerin (BCG) vaccination, incorrect method of TST administration, incorrect interpretation of reaction, and incorrect bottle of antigen used. IGRA blood tests or TSTs (but not both) can be used in most situations when testing for TB infection. However, when substituting IGRA blood tests for TSTs, CDC reminds clinicians that the interpretation criteria for IGRA blood tests differ from that of TSTs. Pharmacists should keep in mind that IGRA blood tests are preferred in persons who have received BCG either as a vaccine or for cancer therapy, as well as in individuals from groups that historically have had poor rates of return for TST reading. On the other hand, TSTs are preferred in children younger than 5 years because data are limited on use of IGRA blood tests in this patient population. In settings with low likelihood of TB exposure or infection, clinicians should consult with public and occupational health authorities and consider deferring routine serial testing. In addition, new CDC guidelines state that annual TB testing of health care personnel who do not have latent TB disease is no longer required. CDC reported that the United States saw a total of 9,029 new TB cases in 2018, which is a 0.7% decrease from the 9,094 cases in 2017. Among the 9,029 TB cases, 69.5% occurred in non–U.S.-born individuals, and 29.5% occurred in U.S.-born individuals. Alaska reported the highest incidence of TB in the nation, with 8.5 cases per 100,000 people. However, CDC said that four states—California, Florida, New York, and Texas—accounted for approximately one-half of the reported cases of TB in the country, “as has been the case for over 2 decades.” While the number of TB cases in the United States continues to decrease, the rate of decline has slowed. As such, CDC reported that the goal of eliminating TB in the country (annual incidence of < 1 case per 1 million persons) is unlikely to become a reality in the 21st century without increased investment and effort. Clinicians can monitor the status of the Aplisol shortage at www.fda.gov/vaccines-blood-biologics/safety-availability-biologics/cber-regulated-products-current-shortages.

A comparative study of topical cantharidin and intralesional PPD to treat molluscum contagiosum

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.

Potential of the dual IFN-γ/IL-2 fluorescence-immunospot assay to distinguish different stages in bovine tuberculosis

Human studies have identified the potential of measuring Mycobacterium tuberculosis specific IFN-γ and/or IL-2 secreting T cell subsets to distinguish different clinical stages of human tuberculosis (TB). To assess these functional T cell subsets in different states of bovine TB we have established a bovine dual IFN-γ/IL-2 fluorescence-immunospot (FluoroSpot) assay and analysed the frequencies of Mycobacterium bovis (M. bovis) specific IL-2 and/or IFN-γ producing cells in PBMC from 30 cattle naturally infected with M. bovis. Depending on their post mortem results the animals were grouped in 22 cattle with visible lesions (VL) and 8 cattle without visible lesions (NVL). In response to bovine tuberculin purified protein derivative (PPD-B) the frequencies of cytokine producing cells and proportions of IL-2 single producers were significantly higher in VL compared to NVL while PWM-induced cytokine responses were similar between the two groups. Dual IL-2+IFN-γ+ T cells could be identified as the largest PPD-B responsive T cell subset in both cattle groups. In conclusion, our FluoroSpot is a valid method to enumerate individual antigen-specific IFN-γ+ and IL-2+ T cell subsets ex vivo. The greater levels of single IL-2 producing T cells associated with the presence of pathology could be a potential biomarker for active TB in cattle.

Simple and rapid liquid chromatographic and electrophoretic methods for phenol quantification and its stability in tuberculin purified protein derivative preparations

Phenol is commonly used as an antimicrobial agent with an initial concentration of 0.35% (w/v) in injectable diluted tuberculin purified protein derivative (TPPD) solution. The anti-microbial action of phenol in TPPD is directly concentration dependent. Furthermore, high phenol content (>0.5%) may have a negative effect on the stability and clinical effectiveness of TPPD solution. Therefore, simple, rapid and reliable reversed phase liquid chromatographic (RPLC) and capillary zone electrophoretic (CZE) methods were firstly developed and validated for phenol quantification in Connaught tuberculin (CT68) PPD diluted preparations at 5 TU per test dose of 0.1 mL. In RPLC, the elution was carried out by 80% (v/v) ACN mixed with 20% (v/v) phosphate buffer containing 0.05% (v/v) triflouroacetic acid (pH 3.2) at 0.2 mL min−1 flow rate and 20.0 °C column temperature. In addition, phenol was separated from tuberculin (CT68) protein with a resolution of (R = 2.81) and was quantified within 3 min.In CZE, the migration of phenol was performed by 50 mmol L−1 borate buffer (pH 9.8) at −20 kV applied voltage and 25.0 °C capillary temperature. Furthermore, excellent linearity was achieved within 0.17–0.53% (w/v) for the phenol content with coefficients of determination (r2) higher than 0.9995. Moreover, the detection and quantification limits were found to be 0.046 & 0.153% and 0.051 & 0.171% (w/v) with RPLC and CZE respectively. Additionally, the intraday precision (RSD%, n = 9) was ranged between 0.18 and 0.39 and 0.33–54 with RPLC and CZE respectively. Moreover, the interday precision (RSD%, n = 27) was varied between 2.06 and 2.99 and 2.25–3.40 by RPLC and CZE, respectively. Furthermore, the obtained mean recoveries were ranged between 91.32 and 107.51% with RPLC and 90.71–108.92% with CZE. In addition, the effect of different storage temperatures at 4, 25 and 37 °C over storage periods of 2, 7, 14, 21 and 30 days was also studied on the TPPD product. The obtained results have revealed that the phenol content was effectively decreased about 37% of its original content after 30 days at storage temperatures of 25 and 37 °C. However, the phenol content did not change and was stable up to 21 days at storage temperature of 4 °C. Therefore, the simple and rapid proposed analytical methods could be used for a rapid expiry investigation of TPPD products based on phenol quantification, as a marker.

Evaluation of tuberculin skin test (TST) in vervet monkeys (Chlorocebus pygerythrus) using experimentally tuberculin sensitized animals.

Nowadays, several test methods with different useful values are available for diagnosis of the tuberculosis (TB) in non-human primates (NHPs). Despite some limitations of tuberculin skin test (TST), it is still the most commonly used method for TB testing of NHPs. During this investigation, TST was performed upon three groups of experimentally tuberculin sensitized and one group of non-sensitized vervet monkeys (Chlorocebus pygerythrus) by means of two types of old tuberculin (OT) and two types of purified protein derivative (PPD) tuberculin. The data obtained from this study revealed that PPD tuberculin prepared from both Mycobacterium tuberculosis and Mycobacterium bovis has more advantages over OT in tuberculin testing of the vervet monkeys. The potency of the PPD tuberculin prepared from Mbovis was estimated almost twice as much of the Mtuberculosis. Intrapalpebral injection of 0.1mL of a concentration of ≥1mg/mL of PPD tuberculin prepared from Mbovis is the preferred method for TST of vervet monkeys.

Skin test of tuberculin purified protein derivatives with a dissolving microneedle-array patch.

The tuberculin skin test (TST) is a long-established screening method for tuberculosis. However, the Mantoux technique is often difficult to reliably perform, which affects testing results and safety, which causes local skin pain and pruritus. In this study, dissolving microneedle-array patches (MNP) were used to deliver purified protein derivative (PPD) tuberculin into the skin. The skin reaction was compared between MNP delivery and conventional injection. The MNP penetrated the skin easily with a thumb press, and the microneedle dissolved into the skin completely after 1h. The storage life of MNP loaded with PPD (MNP-PPD) was 7weeks at atmospheric pressure and room temperature. Only 1/50 dosage of PPD (approximately 0.04IU) was needed in MNP compared with conventional injection (2IU) in terms of skin reactivity to TST. When TST was tested in volunteers, the redness and induration of the skin were 19.7 ± 5.6mm in TB patients, 12.6 ± 4.4mm in LTBI (latent TB infection) patients, and 5.8 ± 2.7mm in BCG vaccination healthy volunteers and lasted approximately 26 ± 5.4days. When applied with MNP-PPD, the redness and induration on the skin decreased significantly to 3.1 ± 0.7mm in TB patients and 2.0 ± 0.5mm in LTBI, and the duration time was only 8.5 ± 1.5days. Moreover, despite the relatively mild skin reactivity in BCG vaccination healthy volunteers with conventional injection, there was no skin reactivity in BCG vaccination healthy volunteers with MNP-PPD. In addition to being minimally invasive, needle-free, and painless, no adverse effects were attributed to the new diagnostic method, which may be of value for the safe and effective clinical administration of TB screening. When applied with MNP-PPD, an area of redness and induration greater than 2.5mm can identify a TB-positive patient.

Characteristics of miliary tuberculosis in pregnant women after in vitro fertilisation and embryo transfer.

To analyse the clinical characteristics of miliary tuberculosis (TB) in pregnant women after in vitro fertilisation and embryo transfer (IVF-ET). Six pregnant women with miliary TB after IVF-ET between October 2010 and July 2015 were retrospectively analysed. The patients were aged between 29 and 39 years. Fever, cough and dyspnoea were the main symptoms, and started during the first and second trimesters of pregnancy. Erythrocyte sedimentation rate and C-reactive protein were increased. White blood cell count was normal or slightly increased. Tuberculin purified protein derivative and acid-fast smear of sputum tests were negative. T-SPOT®.TB was performed in two cases and the results were positive. Chest computed tomography showed typical miliary nodules. One patient had tuberculous meningitis. Although all patients were cured, the foetuses died. The principal symptoms of miliary TB in pregnant women after IVF-ET were fever and dyspnoea. Physicians should consider the occurrence of miliary TB, particularly in the case of fever of long duration, respiratory symptoms and no response to antibiotic treatment.

Efficacy of tuberculin purified protein derivative (PPD) immunotherapy in the treatment of plane warts on face

Efficacy of tuberculin purified protein derivative (PPD) immunotherapy in the treatment of plane warts on face

A New Model to Calibrate a Reference Standard for Bovine Tuberculin Purified Protein Derivative in the Target Species.

Since 1986, use of a Bovine International Standard (BIS) for bovine tuberculin has been required to ensure national and international uniformity regarding the potency designation of bovine tuberculin Purified Protein Derivative (PPDb) preparations produced by multiple manufacturers. The BIS is the unique golden standard in the guinea pig potency assay, representing 100% potency, where potencies of production batches are calculated as relative potencies in comparison with the potency of the BIS which was set at 32,500 international Unit (IU) per mg. The stock supply and lifetime of the BIS is limited.The aim of this study was to develop a model to determine the potency of a newly produced in-house Reference Standard (RS) for PPDb with great accuracy in the target species (cattle) and to prove its precision and accuracy in the guinea pig potency test. First simulations were done to estimate the required number of cattle needed. Then, 30 naturally bTB infected cattle were subjected to a tuberculin skin test using multiple injections of both the RS and the BIS. Both were applied randomly in the same volume and concentration (1 dose). The potency of the RS against the BIS was directly derived from the least square means (LSMEANS) and was estimated as 1.067 (95% CI: 1.025–1.109), equal to a potency of 34,700 ± 1,400 IU/mg. In six guinea pig potency assays the RS was used to assign potencies to production batches of PPDb. Here, precision and accuracy of the RS was determined according to the parallel-line assay. Relative potencies were estimated by exponentiation of the common slope. The corresponding 95% confidence intervals were obtained according to Fieller's theorem. In sensitized guinea pigs, the relative potency of the RS against the BIS was 1.115 (95% CI: 0.871–1.432), corresponding to an absolute potency of 36,238 IU/mg (95% CI: 28,308–46,540).In conclusion: the method used to determine the potency of the RS against the BIS in naturally bTB infected cattle, resulted in a highly accurate potency estimate of the RS. The RS can be used in the guinea pig test to assign potencies to PPDb production batches with high precision and accuracy.

Altered monocyte phenotypes but not impaired peripheral T cell immunity may explain susceptibility of the elderly to develop tuberculosis.

Tuberculosis (TB) is the leading killer due to a single infectious disease worldwide. With the aging of the global population, the case rate and deaths due to TB are highest in the elderly population. While general immunosenescence associated with old age is thought to contribute to the susceptibility of the elderly to develop active TB disease, very few studies of immune function in elderly individuals with Mycobacterium tuberculosis (M.tb) infection or disease have been performed. In particular, impaired adaptive T cell immunity to M.tb is one proposed mechanism for the elderly's increased susceptibility primarily on the basis of the decreased delayed type hypersensitivity response to tuberculin-purified protein derivative in the skin of elderly individuals. To investigate immunological reasons why the elderly are susceptible to develop active TB disease, we performed a cross-sectional observational study over a five year period (2012-2016) enrolling participants from 2 age groups (adults: 25-44 years; elderly: 65 and older) and 3 M.tb infection statuses (active TB, latent TB infection, and healthy controls without history of M.tb infection). We hypothesized that impaired peripheral T cell immunity plays a role in the biological susceptibility of the elderly to TB. Contrary to our hypothesis, we observed no evidence of impaired M.tb specific T cell frequency or altered production of cytokines implicated in M.tb control (IFN-γ, IL-10) in peripheral blood in the elderly. Instead, we observed alterations in monocyte proportion and phenotype with age and M.tb infection that suggest their potential role in the susceptibility of the elderly to develop active TB. Our results suggest a potential link between the known widespread low-grade systemic inflammation of old age, termed "inflammaging," with the elderly's specific susceptibility to developing active TB. Moreover, our results highlight the need for further research into the biological reasons why the elderly are more susceptible to disease and death from TB, so that public health systems can be better equipped to face the present and future problem of TB in an aging global population.

Prevalence of Bovine Tuberculosis among Buffaloes Innorth Karnataka

The present study was undertaken to assess the prevalence of bovine tuberculosis among buffaloes in Belgaum, Dharwad and Bagalkote districts of North Karnataka, for a period of eight years (2002 to 2009). A total of 2184 buffaloes belonging to 30 organized farms were screened by single intradermal tuberculin test using bovine tuberculin Purified protein derivative. Overall prevalence of tuberculosis among buffaloes was found to be 0.96 per cent. Prevalence was higher in females (0.97%) than males (0.84%) and in Murrah (1.02 %) than Surthi (0.85 %). Analysis of the temporal pattern revealed a gradual declining trend over a period of eight years.

A comparative study between intralesional PPD and vitamin D3 in treatment of viral warts

&lt;p class="abstract"&gt;&lt;strong&gt;Background:&lt;/strong&gt; 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.&lt;/p&gt;&lt;p class="abstract"&gt;&lt;strong&gt;Methods:&lt;/strong&gt; 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.&lt;strong&gt;&lt;/strong&gt;&lt;/p&gt;&lt;p class="abstract"&gt;&lt;strong&gt;Results:&lt;/strong&gt; 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.&lt;/p&gt;&lt;p class="abstract"&gt;&lt;strong&gt;Conclusions:&lt;/strong&gt; Intralesional vitamin D and intralesional Tuberculin PPD immunotherapy shows significant results in treatment of viral warts.&lt;/p&gt;

Surface-modified cellulose paper and its application in infectious disease diagnosis

A sensitive and stable cellulose paper platform modified with carboxymethyl cellulose (CMC) was investigated for use as a rapid and accurate immunoassay for tuberculosis. Absorbing CMC to the paper substrate enables it to act as a linker for immobilizing biomolecules in a hydrophilic environment to promote their functionality and stability. This CMC-modified cellulose material can also be crosslinked with 1-ethyl-3-(-3-dimethylaminopropyl) carbodiimide hydrochloride/N-Hydroxy-succinimide (EDC/NHS) to generate NHS ester groups on the paper surface that can be covalently conjugated with ligands or target analytes for highly specific binding. As a result, with this system we were able to achieve a detection limit of 0.03 ng/mL for tuberculin-purified protein derivative. In addition, multiple tests can be completed simultaneously in as little as 21 min. These results demonstrate how this surface-modified paper analytical device can provide sensitive and high-throughput on-site disease diagnosis for resource-limited settings.

Antigen-dependent effects of divergent selective breeding based on natural antibodies on specific humoral immune responses in chickens

NAb are defined as antigen binding antibodies present without a known previous exposure to this antigen. NAb are suggested to enhance specific antibody (SpAb) responses, but consequences of different NAb levels on immunization are largely unknown. Layer chickens were divergently selected and bred for keyhole limpet hemocyanin (KLH)-binding NAb titers, resulting in a High line and a Low line. In this study, we investigated: (1) the relation of NAb levels with SpAb titers; and (2) the effect of immunization on NAb titers. The 50 highest females of the High line and the 50 lowest females of the Low line of generation 2 were intramuscularly immunized at 33 weeks of age with 1 mL phosphate buffered saline (PBS) containing one of four treatments: (1) negative control (no antigen), (2) 500 µg KLH, (3) 100 µg avian tuberculin purified protein derivative of Mycobacterium avium (PPD), or (4) 250 µg human serum albumin (HuSA). IgM and IgG titers of NAb and SpAb in plasma were determined prior to immunization and weekly for 5 weeks post immunization by indirect ELISA. In addition, antibody affinity was investigated. No differences in SpAb and NAb response against KLH and PPD were observed as a consequence of different NAb titers, but increased and prolonged SpAb and NAb titer responses against HuSA were observed for the High line compared to the Low line. Different natural antibody titers did not impair SpAb dynamics and SpAb affinity. NAb titers were not, or for only short-term, affected by immunization. We show here that NAb may enhance SpAb responses, but that this effect is antigen-dependent. We hypothesize that NAb play a role in general disease resistance through enhancement of the humoral adaptive immune response.

Impaired TNF, IL-1β, and IL-17 production and increased susceptibility to Mycobacterium tuberculosis infection in HTLV-1 infected individuals

IFN-γ and TNF play critical roles in the control of Mycobacterium tuberculosis infection. Despite leading to an exaggerated production of inflammatory cytokines, HTLV-1 infection increases the risk of developing tuberculosis (TB). However, the immune mechanisms accounting for this phenomenon are still unclear. The aim of this study was to evaluate immunological aspects of the HTLV-1/M. tuberculosis co-infection. In this cross-sectional study, the levels of TNF, IL-1β, and IL-17 were determined by ELISA in the supernatants of either unstimulated or tuberculin purified protein derivative (PPD) stimulated peripheral blood mononuclear cells. Cells from HTLV-1 infected individuals produced lower levels of TNF following PPD stimulation compared to unstimulated cells. IL-1β and IL-17 production by cells from HTLV-1/M. tuberculosis co-infected individuals was lower than in cells from patients with TB. Impairment in TNF, IL-1β, and IL-17 production upon stimulation with mycobacterial antigens may contribute to the increased susceptibility to M. tuberculosis infection observed in HTLV-1 infected individuals.