Demonstração do complemento total no liquido cefalorraqueano normal

Reis-Filho, João Baptista

doi:10.1590/S0004-282X1978000100001

Resumos

Até o presente momento, todos os autores afirmam que o líquido cefalorraqueano (LCR) normal do homem não apresenta atividade hemolítica do complemento total. Alguns pesquisadores verificaram, entretanto, a presença de todos os quatro componentes em muitas amostras de LCR normal sem conseguir, porém, demonstrar a atividade hemolítica do complemento total. Este fato parecia indicar que o LCR normal era desprovido da atividade hemolítica do complemento total provavelmente por motivo da concentração insuficiente de suas proteínas. Para investigar esta hipótese foi feita uma prova preliminar, concentrando 10 amostras de LCR normal, dividindo-se o concentrado em duas partes, uma das quais foi inativada à temperatura de 56°C, durante 20 minutos. Esta prova revelou hemólise das hemácias sensibilizadas somente em relação com o LCR concentrado ativo. Em prosseguimento foram feitas pesquisas em 108 amostras de LCR normal obtidas por punção cisternal, selecionadas de pacientes sem doença de ordem geral, porém com perturbações neurológicas ou psíquicas que, em geral, não costumam determinar alterações do LCR. As amostras de LCR foram concentradas vinte vezes o seu volume, pelo método da diálise sob pressão negativa em camisa de colódio, segundo o processo de Mies. Esta pesquisa proporcionou resultados que permitiram concluir que o líquido cefalorraqueano normal concentrado apresenta atividade hemolítica do complemento total em 98% dos casos. O título do complemento total variou de 0 a 23 unidades de hemólise 50% em 1 ml de líquido concentrado.

It has long been known that total complement activity is absent in normal cerebrospinal fluid (CSF). Lewandowsky (1900) was the first to call attention to the fact and this finding has been confirmed by other investigators. There is only the discrepant article of Heidelberger & Muller (1949) whose peculiar results may be questioned, because what they observed in their experiments was the activating effect of CSF over serum complement, never an increase of the amount of complement itself. In 1922, Banchieri, Göckel and Kafka demonstrated that the first component of complement (mid-piece) is always present in normal CSF and the studies in this line of investigation have progressed and other complement components were shown to be present in it. Kuwert et al. (1964) have demonstrated the presence of the first component in all CSF specimens, the fourth and the second in the majority of the specimens, and the third component in approximately 30 per cent of the cases. However, in spite of the presence of the four components in many CSF specimens, these authors were not able to demonstrate total complement activity in any case. This puzzling abnormality led Kuwert et al. to assume that normal CSF complement system is defective. Considering this hard problem carefully, the idea occurred to us that the low protein content would account for absence of total complement activity in normal CSF. In order to investigate this hypothesis, tests for complement activity were performed in a series of 108 normal CSF specimens. These specimens were routinely examined for cell count, proteins, sugar and chloride contents, globulins tests, complement fixation tests for syphilis and cysticercosis and all specimens were within normal values. Specimens containing blood were discarded. All specimens were concentrated in order to get a 20 times enrichment of the CSF proteins, according to Mies method of dialysis in collodion bag under negative pressure of 400 mm Hg. The tests for complement activity were performed as soon as possible (usually within four hours) after the withdrawal of CSF from the patients. For the demonstration of total complement activity the quantitative method of Wadsworth et al. (1931), based on photometric determination of the 50 per cent hemolytic endpoint was used. The titer is indicated by number of complement units in 1 ml of concentrated CSF. The results of this investigation have demonstrated total complement activity in 106 normal CSF specimens. Up to now, to the best of our knowledge this technique of investigation and the demonstration of total complement activity in normal CSF have not been previously described in the literature. The following conclusions can be drawn from this study: 1 — when concentrated 20 times, normal cisternal cerebrospinal fluid showed the presence of total complement activity in 98 per cent of the cases; 2 — the complement titer ranged from 0 to 23 units in 1 ml of the concentrated fluid; 3 — the statement that normal cerebrospinal fluid complement system is defective must be reconsidered; 4 — the results of this investigation open a new field of research in cerebrospinal fluid physiology and pathophysiology.

João Baptista Reis-Filho

RESUMO

Até o presente momento, todos os autores afirmam que o líquido cefalorraqueano (LCR) normal do homem não apresenta atividade hemolítica do complemento total. Alguns pesquisadores verificaram, entretanto, a presença de todos os quatro componentes em muitas amostras de LCR normal sem conseguir, porém, demonstrar a atividade hemolítica do complemento total. Este fato parecia indicar que o LCR normal era desprovido da atividade hemolítica do complemento total provavelmente por motivo da concentração insuficiente de suas proteínas. Para investigar esta hipótese foi feita uma prova preliminar, concentrando 10 amostras de LCR normal, dividindo-se o concentrado em duas partes, uma das quais foi inativada à temperatura de 56°C, durante 20 minutos. Esta prova revelou hemólise das hemácias sensibilizadas somente em relação com o LCR concentrado ativo. Em prosseguimento foram feitas pesquisas em 108 amostras de LCR normal obtidas por punção cisternal, selecionadas de pacientes sem doença de ordem geral, porém com perturbações neurológicas ou psíquicas que, em geral, não costumam determinar alterações do LCR. As amostras de LCR foram concentradas vinte vezes o seu volume, pelo método da diálise sob pressão negativa em camisa de colódio, segundo o processo de Mies. Esta pesquisa proporcionou resultados que permitiram concluir que o líquido cefalorraqueano normal concentrado apresenta atividade hemolítica do complemento total em 98% dos casos. O título do complemento total variou de 0 a 23 unidades de hemólise 50% em 1 ml de líquido concentrado.

SUMMARY

It has long been known that total complement activity is absent in normal cerebrospinal fluid (CSF). Lewandowsky (1900) was the first to call attention to the fact and this finding has been confirmed by other investigators. There is only the discrepant article of Heidelberger & Muller (1949) whose peculiar results may be questioned, because what they observed in their experiments was the activating effect of CSF over serum complement, never an increase of the amount of complement itself. In 1922, Banchieri, Göckel and Kafka demonstrated that the first component of complement (mid-piece) is always present in normal CSF and the studies in this line of investigation have progressed and other complement components were shown to be present in it. Kuwert et al. (1964) have demonstrated the presence of the first component in all CSF specimens, the fourth and the second in the majority of the specimens, and the third component in approximately 30 per cent of the cases. However, in spite of the presence of the four components in many CSF specimens, these authors were not able to demonstrate total complement activity in any case. This puzzling abnormality led Kuwert et al. to assume that normal CSF complement system is defective. Considering this hard problem carefully, the idea occurred to us that the low protein content would account for absence of total complement activity in normal CSF. In order to investigate this hypothesis, tests for complement activity were performed in a series of 108 normal CSF specimens. These specimens were routinely examined for cell count, proteins, sugar and chloride contents, globulins tests, complement fixation tests for syphilis and cysticercosis and all specimens were within normal values. Specimens containing blood were discarded. All specimens were concentrated in order to get a 20 times enrichment of the CSF proteins, according to Mies method of dialysis in collodion bag under negative pressure of 400 mm Hg. The tests for complement activity were performed as soon as possible (usually within four hours) after the withdrawal of CSF from the patients. For the demonstration of total complement activity the quantitative method of Wadsworth et al. (1931), based on photometric determination of the 50 per cent hemolytic endpoint was used. The titer is indicated by number of complement units in 1 ml of concentrated CSF. The results of this investigation have demonstrated total complement activity in 106 normal CSF specimens. Up to now, to the best of our knowledge this technique of investigation and the demonstration of total complement activity in normal CSF have not been previously described in the literature. The following conclusions can be drawn from this study: 1 — when concentrated 20 times, normal cisternal cerebrospinal fluid showed the presence of total complement activity in 98 per cent of the cases; 2 — the complement titer ranged from 0 to 23 units in 1 ml of the concentrated fluid; 3 — the statement that normal cerebrospinal fluid complement system is defective must be reconsidered; 4 — the results of this investigation open a new field of research in cerebrospinal fluid physiology and pathophysiology.

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Demonstração do complemento total no liquido cefalorraqueano normal

Demonstration of total complement activity in normal cerebrospinal fluid

Datas de Publicação

Publicação nesta coleção
28 Ago 2012
Data do Fascículo
Mar 1978

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[1] 1. ALPER, C. A.; PROPP, R. P.; KLEMPERER, M. R. & ROSEN, F. S. Inherited deficiency of the third component of human complement (C'3). J. clin. Invest. 48:553,1969.

[2] 2. ASHERSON, G. L. Serum complement levels in systemic lupus erythematosus and other diseases. Aust. Ann. Med. 9:57, 1960.

[3] 3. AUDRAN, R. Notions actuelles concernant le complément. Rev. Europ. Étud. Clin. Biol. 15:610, 1970.

[4] 4. AUSTEN, K. F.; BECKER, E. L.; BORSOS, T.; LACHMANN, P. J.; LEPOW, I. H.; MAYER, M. M.; MÜLLER-EBERHARD, H. J.; NELSON, R. R.; RAPP, H. J.; ROSEN, F. S. & TRNKA, Z. Nomenclature of complement. Immuno-chemistry 7:111, 1970.

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