Tuberculin PPD Research Articles

Lymphocyte unresponsiveness to PPD tuberculin in badgers infected with Mycobacterium bovis

Lymphocyte transformation tests using phytohaemagglutinin and concanavalin A showed that both T and B lymphocytes are functional in tuberculous badgers. Dose response curves indicated that purified protein derivative sensitised lymphocytes were suppressed when cultured in the presence of the antigen.

Solitary pulmonary nodule in sarcoidosis

ROENTGENOGRAPHIC features of pulmonary sarcoidosis include bilateral symmetrical hilar adenopathy, alveolar infiltrates, chronic upperlobe fibrosis and cavitary disease, and reticular or nodular interstitial infiltrates.¹Unusual roentgenographic manifestations of this disease are giant bulla formation, multiple nodules, pleural thickening and effusion, and nonsymmetrical hilar or perihilar masses.²The solitary pulmonary nodule is a rare manifestation of pulmonary sarcoidosis. <h3>Report of a Case</h3> A 32-year-old black woman was in excellent health until she lost weight, and a nonproductive cough, malaise, and lowgrade fever developed. An intermediate-strength PPD tuberculin test had been positive for approximately ten to 15 years, without a history of isoniazid prophylaxis. She smoked two packs of cigarettes per day for approximately 15 years and has a history of sickle cell trait. The patient is from the West Indies and has no history of recent travel to any area of endemic fungal disease. Her vital signs were normal,

Solitary Pulmonary Nodule in Sarcoidosis

ROENTGENOGRAPHIC features of pulmonary sarcoidosis include bilateral symmetrical hilar adenopathy, alveolar infiltrates, chronic upperlobe fibrosis and cavitary disease, and reticular or nodular interstitial infiltrates.¹Unusual roentgenographic manifestations of this disease are giant bulla formation, multiple nodules, pleural thickening and effusion, and nonsymmetrical hilar or perihilar masses.²The solitary pulmonary nodule is a rare manifestation of pulmonary sarcoidosis. <h3>Report of a Case</h3> A 32-year-old black woman was in excellent health until she lost weight, and a nonproductive cough, malaise, and lowgrade fever developed. An intermediate-strength PPD tuberculin test had been positive for approximately ten to 15 years, without a history of isoniazid prophylaxis. She smoked two packs of cigarettes per day for approximately 15 years and has a history of sickle cell trait. The patient is from the West Indies and has no history of recent travel to any area of endemic fungal disease. Her vital signs were normal,

Experimental Infection with Mycobacterium Avium, Serotype 2, in Pigs

Twelve pigs were inoculated orally with Mycobacterium avium. The doses used were 0.5, 2 or 10 mg daily for 5 days, or 10, 50 or 180 mg once (1 mg = 37 × 106 viable units). Two pigs were used per dose, 1 of which was sacrificed 3 days, the other 28/31 days after the last inoculation (Table 1). Three days after inoculation, M. avium was found in the tonsils and in the intestinal mucosa of all 6 pigs, and in the mesenteric lymph nodes of 4. Viable unit counts for tonsils and intestinal mucosa were highest in pigs inoculated with 180 mg×1 and 10 mg×5. Histopathologically these pigs showed activation of the lymphoid tissue in the tonsils, Peyer patches and mesenteric lymph nodes. Twenty-eight/31 days after inoculation a spreading of the infection had taken place in all pigs, most often to the liver, less frequently to the spleen and the lungs. The kidneys and the musculature were not infected (Table 4). A correlation was apparent between the size of dose and the number of viable organisms in the tissues. Divided doses gave about 10 times higher viable counts than a single dose with the same total number of organisms (Table 5). No gross lesions were found 28/31 days after inoculation. Microscopic granulomatous lesions were found in the tonsils of 6 pigs, in the intestinal mucosa of 4 pigs, in the mandibular and mesenteric lymph nodes of 6 pigs, in the retropharyngeal lymph nodes of 3 pigs, and less frequently in the parotid and hepatic lymph nodes (Table 3). Five of 6 pigs were weakly sensitive to avian tuberculin PPD, 1000 t.u. per dose, when tested 22/25 days after inoculation; 1 of these pigs cross-reacted to human tuberculin (Table 2).

PPD-tuberculin and PPD-Battey dual skin testing of hospital employees and medical students.

Tuberculin PPD and PPD-Battey skin tests were simultaneously applied to 3,882 employees of Charity Hospital and 408 medical students at Louisiana State University. The PPD was doubtful (5 to 9 mm induration) in 253 of the total 4,290 persons tested (5.9%). In 86 of these 253 persons, the reaction to PDD-Battey was greater than the reaction to PPD, presumably identifying a subpopulation with a falsely positive PPD and therefore at considerably lower risk of developing future tuberculous disease. Of the 408 medical students (average age 24 years), 80 (19.6%) were classified by the skin tests as having atypical mycobacterial sensitization as compared to six (1.45%) who were classified positive or probably positive to Mycobacterium tuberculosis (P less than .001). In the Southeastern United States, where the incidence of atypical mycobacterial infection is relatively high and occurs at a young age, dual skin testing may have its greatest applicability in identifying tuberculous infection when the PPD falls in the "doubtful" 5 to 9 mm range.

Actinomycosis Presenting as Wegener's Granulomatosis

A MAN with thoracic actinomycosis had disseminated lesions. Initial biopsy specimens showed a granulomatous vasculitis and absent sulfur granules. Several cultures were negative. The correct diagnosis was not made for a long period, and the clinical and pathologic findings suggested a form of Wegener's granulomatosis. It was not until repeated biopsy specimens and cultures showed the organism that the diagnosis of actinomycosis was made. This case illustrates the importance of proper and persistent microbiologic techniques. Report of a Case A 43-year-old man was referred to the hospital because of a six-month history of coughing, weight loss, hemoptysis, coryza, and nosebleeds. At that time, he had a positive PPD tuberculin test, and the chest roentgenogram showed an infiltrate in the right upper lobe and right lower lobe. He also had fluctuant skin masses on the dorsum of the hands bilaterally and on the right thigh. Clinically, these were thought to represent

Production of reference PPD tuberculins for veterinary use in the United States

A method of producing purified protein derivative tuberculin is described which utilizes molecular filtration for the concentrating and washing procedures and a single precipitation of the tuberculo-protein by ammonium sulfate. Information on the relative yields of tuberculo-protein and the levels of impurities in lots of PPD tuberculin prepared from Mycobacterium bovis Strain AN5 and Mycobacterium avium Strain D4 is presented. Reference bovine and avian PPD tuberculins were prepared and evaluated.

Quantification of antigen-reactive cells among human T lymphocytes.

The number of antigen-reactive cells among human peripheral blood T lymphocytes was estimated by a limiting dilution analysis. Antigen-induced lymphocyte activation was measured by means of incorporation of tritiated thymidine [3H]dThd. We have studied the frequency of memory T cells for the bacterial antigens tuberculin PPD and tetanus toxoid in immune donors, as well as the frequency of alloantigen-reactive T cells. In 11 different donors, the observed frequencies of the antigen-reactive T cell ranged between 1:300 and 1:16 000 for PPD; for tetanus toxoid values, between 1:750 and 1:11 500 were obtained in 5 different donors. The frequency of alloantigen-reactive T cells was found to be higher: between 1:200 and 1:600 (n = 10). For 3 donors, the estimated frequencies proved to be reproducible over a period of several months. Finally, a correlation could be demonstrated between the frequency of PPD-reactive T cells and the [3H]dThd incorporation of 4 X 10(4) PPD-stimulated lymphocytes.

Human transfer factor in vitro: identification of L-serine and glycine as components with augmenting effect in lymphocyte transformation assay.

The components of human leucocyte dialysate thought to contain transfer factor, which irresponsible for its capacity to augment antigen or mitogen induced lymphocyte blasogenesis in vitro, were determined by comparing the activity of the original dialysate with its fractions derived from chromatography on Sephadex G‐10. Augmentation of leucoagglutinin (LA) induced transformation was only observed using the original dialysate and two fractions (IIb and IIc) eluting before the total volume of the column. These fractions contained the majority of free amino acids in the dialysate, and no trichloroacetic acid (TCA) precipitable material. Synthetic mixtures, containing the same amount of free amino acids as the fractions, augmented both tuberculin PPD and LA induced blastogenesis in a similar manner to the fractions. The augmenting activity of the synthetic mixture and hence, presumably, the dialysate fractions seemed to be due mostly to l‐serine and glycine, non‐essential amino acids not present in MEM‐S, the culture medium used. No augmenting affect by leucocyte dialysate was seen when MEM‐S was replaced by RPMI 1640, a culture medium containing both l‐serine and glycine. The results suggest that the non‐specific in vitro activity of leucocyte dialysate in this test may be due to a medium supplementation effect and that this phenomenon is probably not relevant to the nonspecific consequences of injection of transfer factor in vivo.

Experimental Infection with Mycobacterium Avium, Serotype 2, in Pigs

Twelve pigs were inoculated orally with Mycobacterium avium. The doses used were 0.5, 2 or 10 mg daily for 5 days, or 10, 50 or 180 mg once (1 mg = 37 × 106 viable units). Two pigs were used per dose, 1 of which was sacrificed 3 days, the other 28/31 days after the last inoculation (Table 1). Three days after inoculation, M. avium was found in the tonsils and in the intestinal mucosa of all 6 pigs, and in the mesenteric lymph nodes of 4. Viable unit counts for tonsils and intestinal mucosa were highest in pigs inoculated with 180 mg×1 and 10 mg×5. Histopathologically these pigs showed activation of the lymphoid tissue in the tonsils, Peyer patches and mesenteric lymph nodes. Twenty-eight/31 days after inoculation a spreading of the infection had taken place in all pigs, most often to the liver, less frequently to the spleen and the lungs. The kidneys and the musculature were not infected (Table 4). A correlation was apparent between the size of dose and the number of viable organisms in the tissues. Divided doses gave about 10 times higher viable counts than a single dose with the same total number of organisms (Table 5). No gross lesions were found 28/31 days after inoculation. Microscopic granulomatous lesions were found in the tonsils of 6 pigs, in the intestinal mucosa of 4 pigs, in the mandibular and mesenteric lymph nodes of 6 pigs, in the retropharyngeal lymph nodes of 3 pigs, and less frequently in the parotid and hepatic lymph nodes (Table 3). Five of 6 pigs were weakly sensitive to avian tuberculin PPD, 1000 t.u. per dose, when tested 22/25 days after inoculation; 1 of these pigs cross-reacted to human tuberculin (Table 2).

Congenital Bronchoesophageal Fistula in the Adult

ESOPHAGEAL - RESPIRATORY tract fistula with esophageal atresia in the infant usually necessitates immediate diagnostic and therapeutic action. Fistulas without atresia often have an insidious clinical course that occasionally persists into adult life. This uncommon cause of chronic pulmonary sepsis needs reemphasis. A recent case prompts this report and an updated review of the literature. Report of a Case A 58-year-old man was admitted to Philadelphia General Hospital in August 1975, with complaints of anorexia, night-sweats without fever, cough, hemoptysis, and weakness in both lower extremities. His past history included chronic alcoholism and a presumptive diagnosis of pulmonary tuberculosis based on positive PPD tuberculin test results and an abnormal infiltrate in the right middle lobe. Sputum cultures were negative for acid-fast bacilli. In 1972, he was treated for three months with isoniazid but subsequently failed to return for followup examination. He also told of numerous unsubstantiated episodes of pneumonia many years

Congenital bronchoesophageal fistula in the adult

ESOPHAGEAL - RESPIRATORY tract fistula with esophageal atresia in the infant usually necessitates immediate diagnostic and therapeutic action. Fistulas without atresia often have an insidious clinical course that occasionally persists into adult life. This uncommon cause of chronic pulmonary sepsis needs reemphasis. A recent case prompts this report and an updated review of the literature. <h3>Report of a Case</h3> A 58-year-old man was admitted to Philadelphia General Hospital in August 1975, with complaints of anorexia, night-sweats without fever, cough, hemoptysis, and weakness in both lower extremities. His past history included chronic alcoholism and a presumptive diagnosis of pulmonary tuberculosis based on positive PPD tuberculin test results and an abnormal infiltrate in the right middle lobe. Sputum cultures were negative for acid-fast bacilli. In 1972, he was treated for three months with isoniazid but subsequently failed to return for followup examination. He also told of numerous unsubstantiated episodes of pneumonia many years

The effect of transfer factor on lymphocyte transformation. Comparison of augmentation by dialysates of leucocytes and lymphoid and non-lymphoid organs.

In this paper we describe experiments to determine whether dialysable extracts of non-lymphoid and lymphoid organs augment lymphocyte transformation in vitro in a manner similar to the augmenting activity of leucocyte dialysates. Human peripheral blood leucocytes were cultured with tuberculin PPD or leucoagglutinin, and dose-related effects of the dialysable extracts on lymphocyte transformation were studied by 125IUdR incorporation. Augmentation of lymphocyte transformation was obtained not only with leucocyte dialysates but also with dialysable extracts of lymphoid and non-lymphoid organs (e.g. spleen, liver, kidney, brain). It is concluded that the agent or agents present in dialysable leucocyte transfer factor preparations, which augment lymphoid transformation in vitro, are widely distributed throughout mammalian tissues.

Regulation of lymphocyte proliferation by soluble products released by stimulated human lymphocytes.

Human lymphocytes pulsed with phytothaemagglutinin (PHA) release mitogenic factors and other lymphokines upon subsequent incubation in fresh medium. Such active supernatants (SUPs) were assayed for their capacity to modify the DNA synthetic response of human lymphocytes exposed to various mitogenic stimuli. Both non-fractionated peripheral lymphocytes and preparations of fractionated T cells were stimulated by active SUPs to DNA synthesis. In contrast, active SUPs reduced the responses of lymphocytes to allogeneic cells, PPD tuberculin and PHA. These reductions were most evident at the end of the culture periods. The results suggest that soluble products released by stimulated lymphocytes activate suppressor lymphocytes which inhibit proliferative responses of lymphocytes.

Infektionsforsøg på svin med Mycobacterium avium, Serotype 2. 2. Peroral infektion med store doser af M. avium

Twelve pigs were inoculated orally with Mycobacterium avium. The doses used were 0.5, 2 or 10 mg daily for 5 days, or 10, 50 or 180 mg once (1 mg = 37 × 106 viable units). Two pigs were used per dose, 1 of which was sacrificed 3 days, the other 28/31 days after the last inoculation (Table 1). Three days after inoculation, M. avium was found in the tonsils and in the intestinal mucosa of all 6 pigs, and in the mesenteric lymph nodes of 4. Viable unit counts for tonsils and intestinal mucosa were highest in pigs inoculated with 180 mg×1 and 10 mg×5. Histopathologically these pigs showed activation of the lymphoid tissue in the tonsils, Peyer patches and mesenteric lymph nodes. Twenty-eight/31 days after inoculation a spreading of the infection had taken place in all pigs, most often to the liver, less frequently to the spleen and the lungs. The kidneys and the musculature were not infected (Table 4). A correlation was apparent between the size of dose and the number of viable organisms in the tissues. Divided doses gave about 10 times higher viable counts than a single dose with the same total number of organisms (Table 5). No gross lesions were found 28/31 days after inoculation. Microscopic granulomatous lesions were found in the tonsils of 6 pigs, in the intestinal mucosa of 4 pigs, in the mandibular and mesenteric lymph nodes of 6 pigs, in the retropharyngeal lymph nodes of 3 pigs, and less frequently in the parotid and hepatic lymph nodes (Table 3). Five of 6 pigs were weakly sensitive to avian tuberculin PPD, 1000 t.u. per dose, when tested 22/25 days after inoculation; 1 of these pigs cross-reacted to human tuberculin (Table 2).

Comparison of fluorescent antibody tests for tuberculosis and paratuberculosis with antigens coupled to insoluble spheres or taken up by macrophages

Sera of 106 cattle from farms with histories of Mycobacterium johnei infection and sera from 15 human tuberculous patients as well as a number of control sera were examined by means of two different fluorescent antibody tests (FAT) for the occurrence of antibodies against M. johnei and M. tuberculosis respectively. The antigens used were PPD johnin and PPD tuberculin. In the macrophage uptake FAT (MU/FAT) mouse macrophages after phagocytosis of the tuberculins served as the matrix; in the tests performed using the defined antigen substrate spheres (DASS) system, Sepharose beads activated by CNBr were used for the matrix. A good correlation was found between the results of the DASS/FAT and those of the MU/FAT, which is known to be a sensitive and specific test in the diagnosis of Johne's disease in cattle. It is suggested that the FAT, with utilization of the DASS system, might have good prospects for routine examination for antibodies against species of Mycobacterium.

Anergy to tuberculin in beef cattle.

Various aspects associated with anergy to tuberculin were investigated in a total of 75 tuberculous cattle. Forty-six (59%) had single lesions and 32 (41%) had multiple lesions. Forty-three (55.1%) had lesions in the lungs or associated lymph nodes and 23 (29.5%) showed multiple involvement of these tissues. Nine anergic cattle had generalised tuberculosis. Histological examination of lesions from 38 animals showed that the majority were of the progressive type. When bovine PPD tuberculin was used more cattle were anergic to the 0.1 mg than to the 0.2 mg dose and fewer tuberculous cattle were detected with either dose if tests were read at 96 h rather than 72h. Other factors thought to contribute to anergy are discussed with special reference to its relative importance in cattle with pulmonary infection.

Depressed cellular immune response to streptococcal antigens in poststreptococcal glomerulonephritis

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 [ 14C] 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.

The production of heterologous antibodies to the human lymphokine: Leukocyte migration inhibitory factor (LIF)

Human leukocyte migration inhibitory factor (LIF) produced by concanavalin A-stimulated lymphocytes was partially purified by Sephadex G-100 chromatography and immunosorption of protein contaminants. This material was injected into two rabbits, and the IgG-IgA fractions of the resulting antisera (anti-LIF) neutralized LIF induced by antigen (PPD tuberculin) with as equal efficiency as that of LIF induced by mitogen. Anti-LIF activity was neither removed by absorption with control supernatant or normal human serum nor was it suppressed by absorption with lymphocytes or lymphoblasts. On the other hand, antibodies against human lymphoid cells (ALG) did not reduce LIF activity, indicating the difference between anti-LIF and classical ALG. In support of this, anti-LIF, in contrast to ALG, was not cytotoxic to lymphocytes, and it did not inhibit spontaneous T-rosette formation with sheep erythrocytes. In crossed immunoelectrophoresis using a conventional proteinstaining technique, only three precipitates appeared. None of these contained LIF. However, a protein migrating in the prealbumin region appeared to be specific for lymphocyte stimulation. The nature and significance of this product is unknown.

Tuberculin sensitivity of cattle inoculated with atypical mycobacteria isolated from cattle, feral pigs and trought water.

Each of 12 cattle was inoculated either subcutaneously and intradermally or into a mesenteric lymph node with 1 of 8 species of liver atypical mycobacteria isolated from cattle, cattle trough water and feral pigs. Seventy-eight days after inoculation the cattle were tuberculin tested with bovine PPD, avian PPD and homologous heat-concentrated syntheic medium tuberculins. They were killed 85 days after inoculation. Organisms were cultured from caseous granulomas at all sites in cattle inoculated with M. avium serotype 2. M. simiae was recovered from a granuloma at the subcutaneous site. Acid-fast bacilli were isolated from the mesenteric lymph node inoculated with trough water organisms. At 72 h, all the cattle had produced skin reactions of 4 mm or more to the homologous tuberculins and all except 1 produced a similar response to avian PPD. Only isolates of bovine origin sensitised cattle to bovine PPD to this degree, and these reactions were less than the corresponding response to avian PPD.