Tuberculin Protein Research Articles

Tuberculosis pericarditis: a case report

Tuberculosis (TB) is still prevalent in the world. In 2020, 1.5 million people died from TB.18 TB accounts for 4% of acute pericarditis. Tuberculous pericarditis (TBP) is due to hypersensitivity to tuberculin protein produced by Mycobacterium tuberculosis and develops in 1-2% of pulmonary TB cases, representing about 1-2% of extrapulmonary tuberculosis. Complications occur in the form of acute pericarditis (4%) and cardiac tamponade (7%), which may require life-saving invasive procedures. Tuberculous pericarditis has an overall mortality rate of 1.43 per 100 person-months over a median follow-up of 11.97 months. Risk factors include diabetes, substance use disorder, HIV-positivity, renal insufficiency, biological or immunosuppressive therapy, and exposure to regions with a high prevalence of tuberculosis. We present a 49-year-old diabetic male with a large complex pericardial effusion occurring three months after a diagnosis of TB pleurisy on antituberculosis drugs, which required pericardiocentesis and pericardial window in the context of cardiac tamponade. Cases of TB pericarditis in developed countries are rarely reported, and there are even fewer in the US. Along with a literature review of the road map for its accurate diagnosis and treatment, this case highlights the relevance of TB in the differential of pericarditis worldwide, even in developed countries.

Efficacy and Safety Profile of Tuberculin Protein Purified Derivative Injection As Immunotherapy For the Treatment of Cutaneous and Anogenital Warts: A Review Article.

Various treatments available today for anogenital and cutaneous warts have limitations, including time-consuming, challenging to perform, and the risk of scarring. A new treatment using tuberculin purified protein derivative (PPD) has been developed, which is expected to generate cellular immunity against HPV. To assess the evidence for the efficacy and safety of PPD treatment for cutaneous and anogenital warts. A literature search was performed with the keyword-based search on digital libraries, including the National Library of Medicine, Cochrane Controlled Register of Trial, and Google Scholar, using the following terms: anogenital warts, condyloma acuminata, cutaneous warts, human papillomavirus, immunotherapy, and tuberculin purified protein derivative. Original studies on treating cutaneous or anogenital warts with PPD were included. The results were 47 clinical trials and 4 case reports. Most of the research was done in countries with common Mycobacterium tuberculosis infection. The treatment showed good efficacy. Comparative studies showed that the treatment has similar efficacy with other immunotherapies. No significant side effects were reported, with evidence of the safety use on the pregnant population. Based on good efficacy and safety, PPD can be considered an alternative therapy, especially in countries where tuberculosis is frequent.

Comparative study of intralesional tuberculin protein purified derivative (PPD) and intralesional measles, mumps, rubella (MMR) vaccine for multiple resistant warts.

Cutaneous warts are frequent conditions that possess much challenge to treat. To evaluate safety and efficacy of intralesional injection of tuberculin purified protein derivative (PPD) antigen injection vs MMR (mumps, measles, rubella) antigen for the treatment of multiple warts. The study included 90 Patients who were divided into three groups (A, B, and C). Each group consisted of (30) patients. Group (A) subjects received intralesional PPD injections. Group (B) subjects received intralesional MMR antigen injection and Group (C) received intralesional saline injection. Full clearance of warts was observed in 18 (60%) of patients in group (A) (70%) vs 24 patients (80%) in group (B). Distal warts completely disappeared in 18 (60%) of patients in group A and 12 (40%) of patients in group B. Infrequent side effects including pain, erythema, and minimal induration were reported in both groups. We established a good safety and efficacy profile for tuberculin PPD and MMR antigens in resistant wart treatment.

Production of monoclonal antibodies to immunogenic peptides of ω-glaidin/c-hordein and rye-secalin for gluten free food screening

Introduction: The ω-glaidin/c-hordein (QPFPQPEQPFPW) and rye-secalin (QPFPQPQQPIPQ) peptides have previously demonstrated immunogenicity in sensitised coeliac T cell lines. The development of monoclonal antibody to those immunogenic peptides is presented with a view of developing an improved enzyme linked immunosorbent assay (ELISA) as a reliable tool to screen the safety of foods specialised for coeliac disease (CD) patients. Methods: Balb/C mice were fed with gluten free food. The immunogens were conjugated to purified tuberculin protein derivative (PPD) with glutaraldehyde and emulsified in Freund’s adjuvant. The employed immunization schedule included 3-5 weeks intervals, followed by a final intravenous injection without adjuvant, 3-4 days prior to fusion. Results: The antibody produced was IgM rather than IgG, although the hybridoma was successfully generated. The IgM class antibody is known to be relatively unstable, so could not be used in kits designed for food screening. Conclusions: The immunogenic peptides could possibly be used to raise monoclonal antibodies for gluten screening. However, a single booster injection might be insufficient to stimulate the spleen directly or the mice should be immunized with higher concentration of immunogen. This could be improved by including multiple booster administrations and increasing the dose.

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.

Immunogenicity of a Recombinant <i>Mycobacterium smegmatis</i> Vaccine Expressing the Fusion Protein CMX in Cattle from Goiás State, Brazil

ABSTRACTThis study aimed to evaluate the immunogenicity of a recombinant Mycobacterium smegmatis vaccine expressing the CMX fusion protein composed of immunodominant epitopes Ag85C, MPT51 and HspX of Mycobacterium tuberculosis, which are important mycobacteria virulence factors. A group of Nelore heifers that were 10 to 12 months of age and negative for the tuberculin skin test (TST) were immunized with four doses of the recombinant vaccine mc2-CMX (M. smegmatis-Ag85C-MPT51-HspX) during a period of one year. Before each immunization, blood was collected to obtain sera for antibody analysis. Serological analysis demonstrated that mc2-CMX was able to induce a humoral response with increased levels of specific IgG antibodies against CMX, despite minimum antibody levels being detected for individual Ag85C, MPT51 or HspX recombinant antigens. However, there was no significant increase in specific CD4+ IFN-γ-positive T cells. Lymphadenomegaly was observed in superficial cervical lymph nodes adjacent to the site of vaccination among mc2-CMX-vaccinated bovines, and the histopathological analysis demonstrated follicular hyperplasia without inflammatory infiltrate or granuloma formation. Animals remained negative for the TST until the end of the experiments, showing no cross-reactivity with the recombinant vaccine and tuberculin proteins. We discuss the potential of mc2-CMX to induce an immune response in cattle.

Intradermal injection of PPD as a novel approach of immunotherapy in anogenital warts in pregnant women

Immunotherapy for treatment of recalcitrant warts was used through different modalities including intralesional injection of purified protein derivative (PPD), which is an extract of Mycobacterium tuberculosis, used for testing exposure to tuberculin protein, either from a previous vaccination or from the environment. This method is used to evaluate the efficacy of a new approach of intradermal injection of PPD in the treatment of anogenital warts in pregnant women. A total of 40 pregnant women, aged 20-35 years, and presented with anogenital warts were enrolled in this study. Human papillomavirus (HPV) typing was done using the GP5+/GP6+ PCR assay. The patients were treated with weekly injections of PPD given intradermally in the forearms, and evaluated for the response regularly. HPV type-6 was the predominant genotype (67.5%). Overall, the improvement in this study was 85% and was related to the extent of tuberculin reactivity. Nineteen (47.5%) patients demonstrated complete clearance, 15 (37.5%) had partial response, and three (7.5%) had minimal response. Three (7.5%) cases did not respond to treatment. Side effects were minimal and insignificant. Treatment of anogenital warts in pregnant women with intradermal injection of PPD was found to be a unique, safe, and effective modality of immunotherapy.

Immunotherapy with Mycobacterium vaccae and Peripheral Blood Flow in Long-treated Leprosy Patients, a Randomised, Placebo-controlled Trial

Objective: to evaluate immunotherapy as a means of improving peripheral blood flow in chronic leprosy patients.Design: this was a double-blind, randomised, placebo-controlled, clinical trial.Materials: heat-killed Mycobacterium vaccae 1mg plus 0.02μg Tuberculin protein per 0.1ml dose in borate buffer, with saline as placebo. Those studied were 92 long-treated residents of a leprosy centre in Iran, 10 of their healthy children and 10 staff members. Evaluation employed the Perimed PF2, Laser–Doppler Flowmeter, a platinum skin thermistor, and a thermal sensibility tester.Methods: single intradermal injections of test or placebo were given to 103 patients 18 months before the blinded evaluation. Fingerpulp blood flux was measured in controlled conditions and vasomotor reflexes and skin sensation to touch, pain and heat were evaluated in 45 and 47 patients in the placebo and M. vaccae groups, respectively, and in 20 healthy control persons.Results: Laser–Doppler flux, skin temperature, vasomotor reflexes and sensation were impaired in leprosy patients. Immunotherapy improved (p < 0.05) Laser–Doppler flux, skin temperature and temperature sensation.Conclusions: immunotherapy, given 18 months earlier, significantly improved blood flow and temperature sensation, in fully-treated, chronic, leprosy patients. The same principles might be employed in other conditions of reduced peripheral blood flow.

An immunochromatographic serological assay for the diagnosis of Mycobacterium tuberculosis

A rapid serological test for tuberculosis (TB) infection was designed using antigens specific to Mycobacterium tuberculosis. Tuberculosis infection, TB vaccination and exposure to environmental Mycobacteria cannot be distinguished using skin tests based on tuberculin protein derivatives. The standard diagnostic techniques such as skin tests, X-rays and DNA techniques are time consuming, expensive, and not practical for screening large populations. We used the 38, 63, 64, 14, 59-kDa antigens of M. tuberculosis to develop a rapid immunochromatographic test kit. This study evaluates the diagnostic potential of the rapid test kit using TB positive and TB negative serum samples from various hospitals in India. The samples were obtained from patients infected with or exposed to bacteria and viral pathogens. The results demonstrated that the combination of antigens improved the diagnostic specificity and sensitivity. The specificity of the test was 99.42% with sensitivity of 98.52% ( n=241). In case of multiple infections, the specificity was 93.15% with a low sensitivity of 73.52% ( n=141). The test kit may offer an improved alternative to purified protein derivative (PPD). This rapid TB test kit may be a useful tool for first-line testing of suspected cases, epidemiological studies and in designing a quality health system to reduce health hazards in resource-poor countries.

Monocyte chemotaxis and chemotactic cytokine release after exposure to hydroxyethyl starch.

Hydroxyethyl starch (HES) is commonly used in leukapheresis and infused as an alternative to blood components for the treatment of hypotension due to hemorrhage and trauma. Its prolonged intravascular persistence and retention in tissue raise concerns about possible effects on humoral and cell-mediated immunity and white cell (WBC) locomotion, particularly in volunteer WBC donors or in severely burned individuals with immunologic depression and increased risk for infection. This study evaluated the effect of HES on human monocyte migration and chemotaxis and the production of antigen- and mitogen-induced WBC-derived chemotactic cytokine. A bioassay was developed to quantitate the neutrophil chemotactic activity of a cytokine generated by mononuclear WBCs stimulated in vitro by phytohemagglutinin or tuberculin protein. The time- and dose-dependent generation of the chemotactic cytokine was not affected by the presence of HES. HES by itself did not induce the generation of this cytokine, nor were human monocyte chemotaxis and spontaneous migration significantly changed by exposure to HES. These results, with those of other investigators, suggest that HES is a safe red cell-sedimenting agent for leukapheresis and an alternative to the use of blood components in shock resuscitation.

Modulation of type I immediate and type IV delayed immunoreactivity using direct suggestion and guided imagery during hypnosis.

Cutaneous reactivity against histamine skin prick test (Type I) and purified tuberculin protein derivative (Mantoux reaction, Type IV) was studied in eight volunteers under hypnosis. Types I and IV immunoreactivity were modulated by direct suggestion (Type I) and guided imagery (Type IV). The volunteers were highly susceptible subjects, selected by means of the Harvard Group Scale of Hypnotic Susceptibility, Form A. When the volunteers underwent hypnotic suggestion to decrease the cutaneous reaction to histamine prick test, a significant (P less than 0.02) reduction of the flare reaction (area of erythema) was observed compared with control histamine skin prick tests. The wheal reaction did not respond to hypnotic suggestion. Neither wheal nor flare reaction could be increased in size by hypnotic suggestion compared with control histamine skin prick tests. A hypnotic suggestion of increasing the Type IV reaction on one arm and decreasing the reaction on the other revealed a significant difference in both erythema size (P less than 0.02) and palpable induration (P less than 0.01). In two cases the reactions were monitored by laser doppler blood flowmetry and skin thickness measurement by ultrasound. The difference between the suggested increased and decreased reaction was 19% for the laser doppler bloodflow (in favor of the augmented side), and 44% for the dermal infiltrate thickness. This study objectively supports the numerous uncontrolled case reports of modulation of immunoreactivity in allergic diseases involving both Type I and Type IV skin reactions following hypnotic suggestions.

Susceptibility of two colonies of Wistar rats to inflammation, with particular reference to delayed hypersensitivity.

Two colonies of Wistar rats were tested for their abilities to produce delayed hypersensitivity reactions and other forms of inflammation. Tuck rats, which respond to dextran with an anaphylactoid reaction, produced delayed reactions to tuberculin protein and to ovalbumin in Freund's incomplete adjuvant. On the other hand, NELP rats which do not respond to dextran also produced delayed reactions to tuberculin protein, but only to ovalbumin when this was contained in Freund's complete adjuvant. Rats of both colonies responded to cotton pellet-induced inflammation, but the adult NELP rats showed resistance both to adjuvant-induced and to collagen-induced arthritis, as well as to the production of experimental allergic encephalomyelitis. NELP rats also showed a much greater reticulo-endothelial system phagocytic activity although antibody titres to sheep red blood cells and the mitogenic activity of concanavalin A and of lipopolysaccharide on spleen cells were similar in the two colonies of rats.

Experimental delayed-onset pulmonary sensitivity: Identification of retest reactions in the lung

An animal model for delayed-onset pulmonary sensitivity has been described ( M. H. Karol, J. Stadler, D. Underhill, and Y. Alarie, 1981, Toxicol. Appl. Pharmacol., 61, 277–285) in which guinea pigs were sensitized by injection with Freund's complete adjuvant. Bronchial provocation challenge (BPC) with tuberculin protein (PPD) 4 to 5 weeks following injection resulted in respiratory reactions with maximal intensity 9 to 12 hr following challenge. Animals were rechallenged bimonthly with PPD aerosol. Responses to second and subsequent inhalation challenges had both earlier onset and earlier wane of response. Retest reactions were maximal 4 to 5 hr following challenge. Intensity of response to each of four challenges remained the same. Histologically, eosinophils were prominent in lungs after repeated BPC. The pulmonary response to repeated inhalation challenge resembled the “retest” reaction noted previously only in the skin. The mechanism of immunologic response appeared to be cell mediated since antibodies were not detected at the height of response, whereas mononuclear cells were abundant in lung tissue. Recognition of a retest reaction in the lung may clarify the pathogenesis of allergic pulmonary reactions occurring in industrial workers 3 to 6 hr following exposure.

Monitoring delayed-onset pulmonary hypersensitivity in guinea pigs

To assess the total pulmonary sensitizing potency of an industrial chemical, consideration must be given to its ability to induce both immediate-onset and delayed-onset respiratory hypersensitivity reactions. An animal model system to evaluate immediate-onset responses has been described (Karol et al., 1980). Measurement of delayed-onset sensitivity in animals may require continuous 48-hr monitoring of respirations. For this purpose a barometric chamber was employed for plethysmography. Delayed sensitivity was produced in guinea pigs by injection of Freund's complete adjuvant, a material classically used to induce delayed immunologic responses. Inhalation challenge of animals with purified tuberculin protein resulted in increased respiratory rate ( X = 46% increase, p < 0.001) 9–12 hr following challenge. Respirations remained elevated for several hours before returning to preexposure levels. Control animals showed no increase. Histologic examination of pulmonary tissue revealed extensive mononuclear cell infiltration consistent with delayed hypersensitivity reactions. This system for measurement of delayed-onset responses has application for assessment of pulmonary sensitizing properties of industrial chemicals as well as testing the efficacy of drugs used for treatment of hypersensitivity reactions.

Cell-mediated immunity to herpes virus type-1 in patients with recurrent corneal herpes simplex.

The proportion of T lymphocytes in the peripheral blood, their phytohaemagglutinin reactivity, and the leucocyte migration inhibitory effect of purified tuberculin protein were found to be equal in 25 recurrent corneal herpes simplex patients and 17 healthy individuals who had never suffered from clinically manifest herpes simplex virus (HSV) infection. Conversely, the HSV vaccine inhibited the leucocyte migration in the healthy group, but not in the recurrent herpes patients when the clinical symptoms were manifest. In 14 recurrent corneal herpes simplex patients, the examinations were repeated after a 3-month symptom-free period, and in these periods the patients corresponded equally to the healthy group in every respect.

Immunostimulators induce granulocyte/macrophage colony-stimulating activity and block proliferation in a monocyte tumor cell line.

Monocyte tumor cell line PU5-1.8 does not normally produce colony-stimulating activity (CSA) required by granulocyte and macrophage progenitors to proliferate and mature in agar. However, CSA is induced in the culture line by as little as 10 ng/ml endotoxic lipopolysaccharide (LPS), with maximum CSA production and release to the medium between 2 and 3 days of incubation. Derived lipid A, but not alkali-treated LPS, is also active. Induction requires RNA and protein synthesis, but is not blocked by mitomycin C or Colcemid. Other inducers of CSA include Mycobacterium Bacillus Calmette-Guérin, tuberculin protein preparation purified protein derivative, zymosan, and phorbol myristate. All inducing agents are specific inhibitors of the monocyte tumor cell proliferation in vitro. Latex beads, another macrophage-activating agent, are rapidly phagocytosed by PU5-1.8 cells, but neither inhibit growth nor induce CSA.

Cytotoxic action of immune lymphocytes on ?adherent? cells (macrophages) of lymph nodes in an autologous system in hypersensitivity of delayed type

The cytotoxic action of immune lymphocytes on “adherent” cells (macrophages; AC) of lymph glands from guinea pigs with hypersensitivity of delayed type (HDT) to streptococcal antigens and to tuberculin proteins was studied in an autologous system. Death of a considerable number of AC during cultivation of a suspension of lymph gland cells from animals sensitized with a streptococcal culture or Bacillus Calmette-Guerin vaccine (BCG) was demonstrated in the presence only of the specific antigen (thermostable fraction of streptococcus or of tuberculin respectively). The discovery of death of AC in an autologous system can be used as a specific and sensitive test with which to study HDT.

Estimativa da prevalência de infeccções micobacterianas em escolares na Guanabara, através de testes intradérmicos com PPD-Rt 23 E PPD-B

Foram testados 543 escolares do Estado da Guanabara, Brasil, na faixa etária de 12 a 14 anos, quanto à hipersensibilidade tuberculínica ao PPD-Rt 23 e ao PPD-B. Confirmou-se a alta prevalência de sensibilidade às tuberculoproteínas em crianças, obtendo-se 33,65% de reatores fortes ao PPD-Rt 23. Da população estudada 85,63% reagiu ao PPD-B, sendo que 6,26% apresentou reação maior a esta tuberculina que ao PPD-Rt 23, sugerindo uma hipersensibilização pelo bacilo Battey, ou outra microbactéria mais relacionada antigenicamente a esse microrganismo que ao Mycobacterium tuberculosis.

Differential effects of immunosuppressants on lymphocyte function.

In vitro and in vivo parameters of T lymphocyte function were evaluated in guinea pigs following treatment with the "cycle-active" drugs, 6-mercaptopurine (6MP) and methotrexate, and the "non-cycle-active" alkylating agent, cyclophosphamide. Commencing at the time of sensitization to tuberculin protein, animals were treated with an 8 day course of one of the cytotoxic drugs. This regimen either reduced or abolished the cutaneous response to PPD. The two cycle-active drugs inhibited the in vitro lymphoproliferative response to PPD and suppressed the elaboration of migration inhibition factor (MIF) by lymph node cells. However, these agents did not reduce blood lymphocytes, deplete the cellularity of the thymic dependent areas of peripheral tissues, or alter the in vitro response of lymph node cells to the nonspecific mitogen PHA. In contrast, treatment with cyclophosphamide was associated with a reduction in peripheral blood and tissue lymphocytes and impaired responses to PHA by residual lymph node cells. In vitro proliferative responses to PPD were inhibited but the capacity of lymph node cells to elaborate MIF was not suppressed. In addition to their effects on antigen-reactive lymphocytes, all three drugs significantly reduced the number of macrophages in induced peritoneal exudates. With respect to immunosuppressive activities, results of these investigations suggest that the noncycle-active agents affect both intermitotic and dividing T lymphocytes without impairing certain intermitotic functions of residual cells. The cycle-active drugs have a more restricted toxicity limited to those T lymphocytes which have been stimulated to undergo active DNA synthesis by antigenic challenge.

Mechanisms of Immunosuppression: Effects of Cyclophosphamide on Cellular Immunity

Abstract A course of cyclophosphamide administered to guinea pigs after immunization with mycobacteria led to a transient inhibition of cutaneous hypersensitivity to tuberculin protein. This treatment resulted in generalized depletion of lymphoid cells, including lymphopenia, and a substantial reduction in the number of macrophages found in induced peritoneal exudates. Results of in vivo cell transfer studies indicated that cyclophosphamide-treated recipients were rendered transiently unresponsive to purified tuberculin protein (PPD). However, sensitized lymph node cells from drug-treated donors were capable of transferring immunity to normal recipients. In vitro tests of cellular immunity with lymph node cells from drug-treated animals yielded discordant results. The proliferative response to both PHA and PPD was significantly impaired. However, these lymphoid cells functioned normally in assays for migration inhibition factor (MIF). Thus, these data suggest that cyclophosphamide can limit proliferation without impairing intermitotic functions of sensitized lymphocytes, such as the release of MIF. Results of this study suggest that cyclophosphamide does not impair development of a population of specifically sensitized T-type lymphocytes. However, several components of the expression phase are affected; the resultant anergy is probably due to a summation of effects on lymphocytes and macrophages.