Circulating immune complexes in patients with advanced tuberculosis and their association with autoantibodies and reduced CD4 + lymphocytes

We investigated the presence of circulating immune complexes (CICs) in serum from tuberculosis (TB) patients with different degrees of pulmonary involvement. Patients were classified into four groups according to the extent of lung involvement: mild (N = 9), moderate (N = 12), moderate plus (N = 16), and severe cases (N = 10). A search for CICs by the 3.5 percent PEG precipitation test showed that the CIC values of patients with the moderate plus or severe form of pulmonary TB were significantly higher compared to healthy controls and to mild and moderate cases (P<0.01 and P<0.001, respectively). Further analysis demonstrated that increased CIC levels were associated with increased autoantibody production, since this abnormality was more prevalent in patients with advanced disease (P<0.01), who also showed a significant reduction of CD4+ T. lymphocytes. The immunoregulatory and pathogenetic implications of these findings are discussed

Preparation of human rabies vaccine in VERO cell culture using a microcarrier system

1. The rabies virus (Pasteur PV strain) was propagated in VERO cells attached to microcarriers in a 3.7-1 bioreactor. Virus titers of about 10(6) LD50/ml were obtained regularly. 2. Ultrafiltration was efficient for concentrating the virus suspensions, and the sucrose gradient reduced the residual VERO cell DNA to acceptable levels (less than 50 pg/dose). The remaining cell DNA content was evaluated by dot-blot hybridization with a probe prepared with VERO cell DNA. 3. The final virus preparations were inactivated by B-propiolactone treatment, showed a potency higher than 2.5 IU/dose and protected mice experimentally infected intracerebrally with rabies virus (CVS-13.2). 4. This methodology for the production of a rabies vaccine for human use should be of interest to countries where high technology facilities are not available

Imunofluorescencia realizada em cerebros de camundongos infectados com virus rabico-cepa CVS, em diferentes estagios de decomposicao / Immunofluorescence performed in brain of rabies virus infected mice - CVS cepa, at different stages of decomposition

O teste de imunofluorescencia (IF) foi avaliado na deteccao de virus rabico presente em cerebros de carcacas de camundongos infectados com virus da cepa CVS, os quais foram conseguidos atraves de uma combinacao de tratamentos, em que se variaram as temperaturas (4,25 e -20 graus Celsius) e o tempo de armazenamento. No teste de IF realizado com impressoes cerebrais de carcacas que haviam sido submetidas a temperatura de 25 graus Celsius por 12-18h, houve maior dificuldade de visualizacao imediata dos corpusculos de inclusao, enquanto que nos materiais conservados a 4 graus Celsius por ate 48h, as inclusoes foram facilmente reconhecidas. Carcacas mantidas a -20 graus Celsius mantiveram-se viaveis a identificacao pela IF mesmo apos terem sido armazenadas por 720h quando foram feitas as ultimas observacoes. Em carcacas mantidas a 25 graus Celsius por 10h, com tratamento posterior a 4 e -20 graus Celsius, o antigeno rabico nao pode ser identificado atraves da IF, em consequencia da decomposicao das carcacas que ocorrem, respectivamente, apos 10 e 24h. Recomenda-se, portanto, empregar o teste de IF, em carater de rotina, no controle de qualidade da vacina contra a Raiva, no que diz respeito a prova de virus residual (teste de verificacao da inativacao viral), de vez que ele permite esclarecer mortes assintomaticas...

Evaluacion de la calidad de los reactivos que se utilizan en la tecnica de contrainmunoelectroforesis para la determinacion de anticuerpos antirrabicos / Evaluation of the quality of the reagents employed in the counterimmunoelectrophoresis technique for the determination of rabies antibodies

En este estudio se comprobo que el Instituto Butantan produce antigenos y sueros indicadores que se pueden utilizar con exito en la prueba de contrainmunoelectroforesis para titular anticuerpos antirrabicos en personas inmunizadas. No se pudieron demostrar diferencias estadisticamente significativas entre los resultados de las pruebas de estandarizacion realizadas en el Instituto Butantan y las pruebas de control de referencia llevadas a cabo en el Centro Panamericano de Zoonosis. Se propone que el Instituto Butantan produzca y distribuya a nivel nacional los reactivos para que los laboratorios de diagnostico apliquen la tecnica de contrainmunoelectroforesis para la determinacion de anticuerpos antirrabicos.