Immunotherapy with oral, heat-killed, Mycobacterium vaccae in patients with moderate to advanced pulmonary tuberculosis.

A research investigation to evaluate the potential of an oral preparation of Mycobacterium vaccae SRL172 (NCTC 11659) as an immunotherapeutic has been carried out in ten patients with moderate to advanced pulmonary tuberculosis at Carrasco Hospital, Argentina. Comparison was made between oral and injected M.vaccae sharing a mutual control group. Clinical, bacteriological, hematological, radiological and immunological assessments all showed comparable benefits for both injected and oral treatment over those achieved with chemotherapy alone. The only significant difference between results of injected and oral M.vaccae was the failure of the latter to reduce TNF-alpha production by cultured mononuclear cells. A more intensive regime for the oral preparation was used, which as an addition to the directly observed therapy, short-course, treatment should improve results in both drug susceptible and drug-resistant cases. A Phase II Good Clinical Practice trial is now required.

Studies on the activating capacity of heat killed M. tuberculosis and its culture filtrate proteins on polymorphonuclear neutrophils and peripheral mononuclear cells from tuberculosis patients

En este estudio se investiga la eficacia de M. tuberculosis muerto porcalor (Mtbi) y las Proteinas del Filtrado del Cultivo (PFC) en la activación de las células mononucleares (MC) y polimorfonucleares neutrolilos (PMN)de sangre periférica de pacientes tuberculosos. Se evalua en 16 pacientes tuberculosos, HIV- y 12 controles sanos el Estallido Respiratorio, los metabolitos derivados del NO y la producción de IL-2, IL-12 y TNFeÁ por las células estimuladas. Se detecta un incremento en la concentración de TNFeÁ en el sobrenadante de cultivo (s.c.) de PMN al comparar con los valoresbasales y en la evaluada en s.c. de MC y PMN estimulados, al ser comparadas con las del grupo control, excepto para los neutrófilos estimuladoscon PFC. Se mostraron niveles aumentados de IL-12 e IL-2 en s.c. de ambas células, MC y PMN estimuladas por en PTB, mientras que no se hallarondiferencias en los s.c. de los controles. Los valores basales de Estallido Respiratorio (RB) detectada en MC y PMN de pacientes no difirieron significantivamente de los correspondientes al grupo control. La expresión del Estallido Respiratorio en ambos tipos celulares fue menor en los pacientes que en los controles, independientemente del estímulo empleado. Sedeterminaron concentraciones de nitritos más elevadas en los sobrenadantesde las MC estimuladas con Mtbi y PFC provenientes de pacientes, comparadas con las de los controles. Los datos obtenidos relacionados al estímulo de la respuesta celular, nos proporcionan información sobre la inmunidad protectiva contra el M. tuberculosis y, a la vez, aportan algunos recursos útiles para una terapia anti-tuberculosa más eficiente (AU)

The efficacy of heat-killed Mycobacterium tuberculosis (HKMtb) andits culture filtrate proteins (CFP) to activate blood mononuclear cells (MC)and polymorphonuclear neutrophils (PMN) from tuberculosis patientswas investigated. Respiratory burst, NO-derived metabolites, IL-2, IL-12and TNF-¦Á production of stimulated cells from 16 HIV- tuberculosispatients and 12 healthy controls were analyzed. Increased amounts ofTNF-¦Á in supernatants from baseline and stimulated polymorphonuclear and mononuclear cells of tuberculosis patients were detected whencompared with controls, except for CFP stimulated neutrophils. Augmented IL-2 and IL-12 levels were observed in supernatants of both stimulated MC and PMN from TBP while no differences were found in control supernatants. The patients had a lower respiratory burst responsethan the controls, for both cell types, regardless of the stimulus employed. Higher nitrite concentrations were found in HKMtb- and CFP-stimulated mononuclear supernatants from patients, compared with controls. The obtained data of the stimulated cellular responses provides usinformation about the protective immunity against Mycobacterium tuberculosis and some resources to obtain a more efficient anti-tuberculous therapy (AU)1

Potential for immunotherapy with heat-killed Mycobacterium vaccae in respiratory medicine.

Immunotherapy with Mycobacterium vaccae has been shown to be beneficial as part of the treatment for a wide range of diseases. In the respiratory system, the late airway response in bronchial asthma is modified by a single dose and bronchial aspects of hayfever are reduced allowing a major reduction in the use of bronchial dilators. In studies of advanced adenocarcinoma of the lung survival is increased by an average of 4 months when up to five doses of M. vaccae are added to the course of chemotherapy. The quality of life of cancer patients receiving immunotherapy with M. vaccae is improved, even if survival is not increased. It is suggested that the mechanism of action of immunotherapy with heat-killed, borate-buffered M. vaccae is likely to be very similar in all these diseases for which human pulmonary tuberculosis provides a model. In this study, additional immunological data are reported from material stored from an earlier study of immunotherapy for pulmonary tuberculosis to help complete the information on the way that treatment with three monthly injections of heat-killed, borate-buffered M. vaccae (SRL172) may act.

Functional characteristics of neutrophils and mononuclear cells from tuberculosis patients stimulated in vitro with heat killed M. tuberculosis.

BACKGROUND: The major protective immune response against intracellular bacteria, such as Mycobacterium tuberculosis, is a cell-mediated immunity involving neutrophils (PMNs) and peripheral mononuclear cells (MCs), contributing to the clearance of this microorganism and the resolution of the infection. This study was addressed to evaluate PMNs and MCs for their bactericidal function. METHODS: The sample comprised 14 tuberculosis (TB) inpatients (HIV-), and 10 healthy controls (HCo). Peripheral PMNs and MCs were separated by Ficoll-Hypaque and cultured in RPMI with or without heat-killed Mycobacterium tuberculosis (HK Mtb). Respiratory burst (RB), CD11b, IL-8 and TNFalpha receptor expression were assessed by flow cytometry in cells undergoing stimulation or not. Presence of IL-8 and TNFalpha in cell culture supernatants was determined by ELISA. RESULTS: TB patients had a lower RB response than HCo for both cell types (MCs, p <0.05, PMNs, p <0.01) regardless of HK Mtb stimulation. Compared to HCo, PMNs and MCs from TB patients presented a reduced CD11b expression, with the two subject groups showing a decrease in this marker expression following HK. Mtb was added to both cell cultures. Whereas fewer IL-8 and TNFalpha receptors were found when studying MCs and PMNs from TB patients, antigen stimulation significantly raised the expression for both cytokine receptors. Culture supernatants from MCs and PMNs of TB patients contained increased amounts of IL-8 and TNFalpha. CONCLUSIONS: The present findings may provide some explanation as to the different roles played by PMNs and MCs in TB immunopathology.

Immunological consequences of three doses of heat-killed Mycobacterium vaccae in the immunotherapy of tuberculosis.

We report the first study of triple-dose immunotherapy with heat-killed Mycobacterium vaccae (SRL 172) combined with short-course, directly observed chemotherapy in newly diagnosed pulmonary tuberculosis patients. The study was carried out in Rosario, Argentina, where single-dose immunotherapy with M. vaccae has previously been shown effective. Twenty-two HIV seronegative patients, sputum-positive for tubercle bacilli, entered a randomised and partly blinded trial. Twelve patients received injections of SRL 172 and 10 patients received placebo on days 1, 30 and 60 of chemotherapy. All patients were followed up clinically, by sputum bacteriology, chest radiography and haematology. Patients receiving SRL 172 showed faster and more complete clinical improvement, accelerated disappearance of bacilli from sputum, better radiological clearance and a more rapid fall in ESR, than did those receiving placebo. Follow-up continued for a year after therapy and no patient failed treatment or relapsed. Special investigations included longitudinal assessments of respiratory bursts and expression of CD11b on separated polymorphonuclear and mononuclear leukocytes. Tumour necrosis factor alpha (TNF-alpha) was measured in the supernates of cultured cells and both TNF-alpha and interleukin-4 (IL-4) were measured in serum samples. Immunotherapy recipients showed a significantly faster return towards normal values in all the immunological parameters, than did placebo recipients. The results are consistent with a regulatory activity on cellular immunity, reducing the influence of Th2 and enhancing Th1 to the benefit of the patients. This could allow a reduced period of chemotherapy without loss of efficacy and help to prevent the development of multi-drug resistance.

TNF-alpha, TGF-beta and NO relationship in sera from tuberculosis (TB) patients of different severity.

Tuberculosis (TB) is the main cause of death by infection diseases worldwide. Considering that NO, TNF-alpha and TGF-beta participate a great deal in TB immunopathogenesis, we wished to analyse whether these mediators showed some relationship with the degree of pulmonary affectation. The sample comprised 29 TB (HIV-), inpatients with mild-moderate (n = 10) or advanced (n = 19) newly-diagnosed disease, together with 12 healthy controls HCo. Serum nitrite was assessed by reducing nitrate to nitrite, and further measured by the Griess reaction. Levels of TNF-alpha and TGF-beta were determined by ELISA (R&D Systems). Serum levels of TNF-alpha were significantly higher in the advanced TB cases if compared with HCo, (p < 0.05 ) and from values of Mild-Moderate TB patients (p < 0.05). Serum levels of TGF-beta from advanced TB patients have increased values if compared with Hco (p < 0.005) and Mild-Moderate patients (p < 0.05). These values were also significantly different from Mild-Moderate cases + HCo (p = 0.01) Advanced TB patients had significantly reduced nitrite levels compared with those of Mild-Moderate patients and HCo (p < 0.002). Taken as a whole NO-derived metabolites in TB patients (M-M and Advanced cases) remained lower than values in HCo (p = 0.005) A negative correlation was found when comparing the two cytokines with nitrites(r = -0.44 ).TGF-beta and TNF-alpha were positively correlated (r = 0.44, p < 0.01), 0.44, p < 0.01. In synthesis, the inverse correlation found between both cytokines concentrations and NO levels in TB patients may be viewed as a consequence of a more predominant TGF-beta effect.

Aquagenic urticaria: report of a case.

Aquagenic urticaria is a very rare form of physical urticaria induced by contact with water. In this case report, we describe a child with a typical form of the disease in whom other types of physical urticaria were ruled out. Clinical manifestations, investigational methodology, and available treatments were reviewed. Treatment with hydroxyzine, 25 mg daily, was successful after a month follow-up in preventing wheals and erythema. However, mild pruritus is still present after contact with water.

Impaired neutrophil function in patients with pulmonary tuberculosis and its normalization in those undergoing specific treatment, except the HIV-coinfected cases.

Our study investigated whether the respiratory burst (RB) of polymorphonuclear neutrophils from tuberculosis (TB) patients was related with the disease severity or treatment, as well as the circulating levels of TNF-alpha. The sample comprised 57 patients with moderate (n=21) or advanced disease (n=36, 13 of them with HIV coinfection, TB-HIV) and 12 controls. Patients were newly diagnosed (n=27) or under treatment (moderate=14, advanced=10, TB-HIV=6). Cytometric analysis showed that untreated patients had a depressed RB in response to Candida albicans, being more pronounced in the advanced group and nearly absent in TB-HIV cases. A recovered RB was observed in treated patients, except for the TB-HIV cases that continued to show a poor response. TNF-alpha serum levels were increased in untreated patients, mostly in the advanced and TB-HIV groups, and showed an inverse and significant correlation with the RB. Disease severity and anti-TB therapy exerted negative and positive influences on the reactive oxygen intermediates production, respectively.