132
TRANSACTIONS OF THEROYALSOCIETY OF TROPICAL MEDICINEANDHYGIENE(1992)86, 132-133
1.8 5
Human cerebral malaria: a-galactosyl antibodies in cerebrospinal fluid
0
:
1.2 0
Balachandran
Ravindranl
and Bidyut
K.
Da&
‘Department of Applied Immunology, Regional Medical Research Centre (ICMR), Bhubaneswar, India; 2Department of Medicine, SCB Medical College, Cuttack, India
Reports in recent years have attributed a role to host immune responsein the pathogenesisof human cerebral malaria (GRAUet al., 1989; KWIATKOWSKIet al., 1990). Following the demonstration of intrathecal synthesis of immunoglobulins in Thai adults with cerebral malaria (CM) (CHAPELet al., 1987), we recently identified and characterized malarial antibodies in cerebrospinal fluid (CSF) of CM casesin India (MITRA et al., 1991). In this communication we provide evidence for the presenceof high levels of cw-galactosyl antibodies (anti-gal) in CSF of adults with CM. This is the first report on the presence of an autoantibody in CSF of human CM cases.There are many reasonsfor studying anti-gal in CSF: (i) anti-gal is a dominant immunoglobulin (Ig) G autoantibody constituting about l-2% of the total IgG in all human sera (GALILI et al., 1987);(ii) serum anti-gal titres are elevated during Plasmodium falciparum infections (RAVINDRANet al., 1989); (iii) a-galactosyldeterminants havebeendemonstrated in some of the parasite glycoproteins (RAMA-
0
1.0
00
0.8
0 0
0
0
E c g v
0
0.6
00 0
z z
0 0
0.4
0.2
0
0
Qp
0
00
8 u ----VW--
iRl ---------w 8
SAMY & REESE, 1986); and (iv) natural autoantibodies
have been estimated in CSF in other neurological disorders with a view to understanding the pathogenesisof the disease(MATSIOTAet al., 1988). Samnlesof CSF were collected from 20 CM cases(mean age 37.7 years, range 17-60), 27 presumptive casesof CM (EM, no demonstrable parasitaemiain peripheral blood but clinically responding to intravenous quinine therapy; mean age 32.4 years, range 1462) and 18 age-matchedcontrols. Details regarding clinical criteria and collection of CSF samples have been described elsewhere (MITRA et al., 1991). Anti-gal titres in 20-fold diluted CSF samoleswere estimatea by cell enzyme-linked
immunoiorbent
assay
(ELISA)using monolayers of rabbit erythrocytes as described earlier (RAVINDRANet al., 1989). Monospecific anti-human IgG or IgM or IgA conjugated to alkaline phosphatasewere used as detecting reagents.The specificity of the antibody reactivity to a-linked galactosewas confirmed by inhibition of the ELISA reaction in the presence of melibiose. An optical density value of the
mean f3 standard deviations obtained with 18 control CSFs was taken as a cut-off point for positivity. About 70% of CM and PCM caseshad signifciant titres of IaG anti-gal in CSF (Figure). However, IgM and IgA titres were not verv hieh. Sianificant titres of anti-nal IaM were found in only 26.jo/o of CM and 21.7% gf NCM cases,and of IgA in 31.5% and 17.3% of CM and NCM casesrespectively. Since no breach of the blood-CSF barrier is known to occur in human CM (WARRELI, et al., 1986) and intrathecal synthesis of Ig has been reported (CHAPELet al., 1987), it has been proposed that either malarial antigen(s) or mitogen(s) could havebeenthe stimulus for the presenceof antibodies in CSF. Raised levels of antigal in CSF tend to indicate possible polyclonal stimulation of lymphocytes in the brain during CM. However, it is not known whether anti-gal titres increasein CSF during uncomplicated P. falciparum malaria. The role, if Address for correspondence: Dr B. Ravindran, Assistant Director, Department of Applied Immunology, Regional Medical Research Centre (ICMR), Chandrasekharpur, Bhubaneswar751 016, India.
0
I/
CM
&B
IP
PCM
C
Figure.IgG a-galactosyl antibodytines (opticaldensity[OD] at 405nm) in CSFin cerebralmalaria(CM), presumptivecasesof cerebralmalaria (PCM),andnormalcontrols(C). The dottedline indicatesthemeanplus threestandarddeviationsof OD valuesin controls.
any, played by CSF anti-gal in pathogenesisof human CM can only be speculative at this stage. Interestingly, anti-gal autoantibodies are found in sera of Old World monkeys, apes and human beings only and not in other mammals(GALILI et al., 1987). New World monkeys, on the other hand, express a-linked galactosedeterrnmants on most of their nucleated cells (GALILI et al.. 1988). Whether this observation has any bearing on the outcome of P. falcipancm infections in New World monkeys remains to be determined-symptoms resembling CM are never seenin New World monkeys infected with P. falciparum, although someof them are highly susceptible and succumbto infection with very high parasitaemia.In this context undertaking further investigations on the role of anti-gal in human cerebral malaria appearsobligatory. We thank Dr S. Mitra for providing someof the CSF samples and Mr P. K. Sahoo for technical assistance. The Regional Medical ResearchCentre, Bhubaneswar is fully supported by funds from the Indian Council of Medical Research,New Delhi. References
Chapel, H. M., Warrell, D. A., Looareesuwan, S., White, N. J., Phillips, R. E., Warrell, M. J., Supaeanta, V. & Tharavaniji, S. (1987). Intrathecal immunoglobulin synthesisin cerebral malaria. Clinical and Experimental Immunology, 67, 524-530. Galili, U., Clark, M. R., Shohet, S. B., Buehler, J. & Macher, B. A. (1987). Evolutionary relationship between the natural and anti-gal antibody and the Gall-3Gal epitope in primates. Proceedings of the National Academy of Sciencesof the USA, 84,1369-1373.
Galili, U., Shohet, S. B., Kobrin, E., Stults, C. L. M. & Macher, B. A. (1988). Man, apes and old world monkeys
133 differ from other mammals in the expressionof o-galactosyl epitopes on nucleated cells. Journal of Biological Chemisty, 263,1755-1762. Grau, G. E., Taylor, T. E., Molyneux, M. E., Wirima, J. J., Vasalli,,I’., Hommel, M. & Lambert, P. H. (1989). Tumour necrosis factor and diseaseseverity in children with falciparum malaria. New England Journal of Medicine, 320, 1586 1591. Kwiatkowski, D., Hill, A. V. S., Sambou, I., Twumasi, I’., Castracane,J., Monogue, K. R., Cerami, A., Brewster, D. R. & Greenwood, B. M. (1990). TNF concentration in fatal cerebral, non-fatal cerebral and uncomplicated Plasmodium falciparum malaria. Lancer, 336, 1201-1204. Matsiota, I’., Balancher, A., Doyon, B., Guilbert, B., Clanet, M., Kouvelas, E. D. & Avrameas, S. (1988). Comparative study of natural autoantibodies in the serum and cerebrospinal fluid of normal individuals and patients with multiple sclerosisand other neurological disease.Annales de’1Znstitut Pasteurlhmunologie, 139,99-108. Mitra, S., Ravindran, B., Das, B. K., Das, R. K., Das, I’. K. & Rath, R. N. (1991). Human cerebral malaria: charac-
terization of malarial antibodies in cerebrospinalfluid. Clinical and Experimental Immunology,86, 19-21. Ramasamy, R. & Reese, R. T. (1986). Terminal galactose residues and the antigenicity of Plasmodium falciparum glycoproteins. Molecular and Biochemical Parasitology, 19, 91101. Ravindran, B., Satapathy, A. K. & Das, M. K. (1989). Naturally occurring anti-galactosyl antibodies in human Plasmodium falciparum infection-a possible role for autoantibodiesin malaria. Immunology Letters, 19, 137-142. Warrell, D. A., Looareesuwan, S., Phillips, R. E., White, N. J., Warrell, M. J., Chapel, H. M. & Tharavanij, S. (1986). Function of the blood-cerebrospinal fluid barrier in human cerebral malaria: rejection of the permeability hypothesis. AmericanJournal of Tropical Medicine and Hygiene, 35, 882889. Received 2 September 1991; accepted for publication 26 September 1991
Announcement
ROYAL
SOCIETY
OF TROPICAL
MEDICINE
AND
I-IYGIENE
The Transactions of the Royal Society of Tropical Medicine and Hygiene is issued bi-monthly (six parts per year) and publishes not only papers presented at Society meetings and symposia but also papers and correspondence submitted from all over the world on every aspect of tropical medicine and health. Supplements on specialist topics are published from time to time. The Transactions is available to Libraries and non-Fellows at an annual subscription of &96. It is dispatched by Accelerated Surface Post at no extra charge. The annual subscription to Fellows is &40. Applications for Fellowship must be made on the correct form, obtainable from the address below. Rate for bona fide students &25. Further
information
from: The Honorary Secretaries, Royal Society of Tropical Medicine and Hygiene, Manson House, 26 Portland Place, London WIN 4EY
TRANSACTIONS OF THEROYALSOCIETY OF TROPICAL MEDICINEANDHYGIENE(1992)86, 132-133
1.8 5
Human cerebral malaria: a-galactosyl antibodies in cerebrospinal fluid
0
:
1.2 0
Balachandran
Ravindranl
and Bidyut
K.
Da&
‘Department of Applied Immunology, Regional Medical Research Centre (ICMR), Bhubaneswar, India; 2Department of Medicine, SCB Medical College, Cuttack, India
Reports in recent years have attributed a role to host immune responsein the pathogenesisof human cerebral malaria (GRAUet al., 1989; KWIATKOWSKIet al., 1990). Following the demonstration of intrathecal synthesis of immunoglobulins in Thai adults with cerebral malaria (CM) (CHAPELet al., 1987), we recently identified and characterized malarial antibodies in cerebrospinal fluid (CSF) of CM casesin India (MITRA et al., 1991). In this communication we provide evidence for the presenceof high levels of cw-galactosyl antibodies (anti-gal) in CSF of adults with CM. This is the first report on the presence of an autoantibody in CSF of human CM cases.There are many reasonsfor studying anti-gal in CSF: (i) anti-gal is a dominant immunoglobulin (Ig) G autoantibody constituting about l-2% of the total IgG in all human sera (GALILI et al., 1987);(ii) serum anti-gal titres are elevated during Plasmodium falciparum infections (RAVINDRANet al., 1989); (iii) a-galactosyldeterminants havebeendemonstrated in some of the parasite glycoproteins (RAMA-
0
1.0
00
0.8
0 0
0
0
E c g v
0
0.6
00 0
z z
0 0
0.4
0.2
0
0
Qp
0
00
8 u ----VW--
iRl ---------w 8
SAMY & REESE, 1986); and (iv) natural autoantibodies
have been estimated in CSF in other neurological disorders with a view to understanding the pathogenesisof the disease(MATSIOTAet al., 1988). Samnlesof CSF were collected from 20 CM cases(mean age 37.7 years, range 17-60), 27 presumptive casesof CM (EM, no demonstrable parasitaemiain peripheral blood but clinically responding to intravenous quinine therapy; mean age 32.4 years, range 1462) and 18 age-matchedcontrols. Details regarding clinical criteria and collection of CSF samples have been described elsewhere (MITRA et al., 1991). Anti-gal titres in 20-fold diluted CSF samoleswere estimatea by cell enzyme-linked
immunoiorbent
assay
(ELISA)using monolayers of rabbit erythrocytes as described earlier (RAVINDRANet al., 1989). Monospecific anti-human IgG or IgM or IgA conjugated to alkaline phosphatasewere used as detecting reagents.The specificity of the antibody reactivity to a-linked galactosewas confirmed by inhibition of the ELISA reaction in the presence of melibiose. An optical density value of the
mean f3 standard deviations obtained with 18 control CSFs was taken as a cut-off point for positivity. About 70% of CM and PCM caseshad signifciant titres of IaG anti-gal in CSF (Figure). However, IgM and IgA titres were not verv hieh. Sianificant titres of anti-nal IaM were found in only 26.jo/o of CM and 21.7% gf NCM cases,and of IgA in 31.5% and 17.3% of CM and NCM casesrespectively. Since no breach of the blood-CSF barrier is known to occur in human CM (WARRELI, et al., 1986) and intrathecal synthesis of Ig has been reported (CHAPELet al., 1987), it has been proposed that either malarial antigen(s) or mitogen(s) could havebeenthe stimulus for the presenceof antibodies in CSF. Raised levels of antigal in CSF tend to indicate possible polyclonal stimulation of lymphocytes in the brain during CM. However, it is not known whether anti-gal titres increasein CSF during uncomplicated P. falciparum malaria. The role, if Address for correspondence: Dr B. Ravindran, Assistant Director, Department of Applied Immunology, Regional Medical Research Centre (ICMR), Chandrasekharpur, Bhubaneswar751 016, India.
0
I/
CM
&B
IP
PCM
C
Figure.IgG a-galactosyl antibodytines (opticaldensity[OD] at 405nm) in CSFin cerebralmalaria(CM), presumptivecasesof cerebralmalaria (PCM),andnormalcontrols(C). The dottedline indicatesthemeanplus threestandarddeviationsof OD valuesin controls.
any, played by CSF anti-gal in pathogenesisof human CM can only be speculative at this stage. Interestingly, anti-gal autoantibodies are found in sera of Old World monkeys, apes and human beings only and not in other mammals(GALILI et al., 1987). New World monkeys, on the other hand, express a-linked galactosedeterrnmants on most of their nucleated cells (GALILI et al.. 1988). Whether this observation has any bearing on the outcome of P. falcipancm infections in New World monkeys remains to be determined-symptoms resembling CM are never seenin New World monkeys infected with P. falciparum, although someof them are highly susceptible and succumbto infection with very high parasitaemia.In this context undertaking further investigations on the role of anti-gal in human cerebral malaria appearsobligatory. We thank Dr S. Mitra for providing someof the CSF samples and Mr P. K. Sahoo for technical assistance. The Regional Medical ResearchCentre, Bhubaneswar is fully supported by funds from the Indian Council of Medical Research,New Delhi. References
Chapel, H. M., Warrell, D. A., Looareesuwan, S., White, N. J., Phillips, R. E., Warrell, M. J., Supaeanta, V. & Tharavaniji, S. (1987). Intrathecal immunoglobulin synthesisin cerebral malaria. Clinical and Experimental Immunology, 67, 524-530. Galili, U., Clark, M. R., Shohet, S. B., Buehler, J. & Macher, B. A. (1987). Evolutionary relationship between the natural and anti-gal antibody and the Gall-3Gal epitope in primates. Proceedings of the National Academy of Sciencesof the USA, 84,1369-1373.
Galili, U., Shohet, S. B., Kobrin, E., Stults, C. L. M. & Macher, B. A. (1988). Man, apes and old world monkeys
133 differ from other mammals in the expressionof o-galactosyl epitopes on nucleated cells. Journal of Biological Chemisty, 263,1755-1762. Grau, G. E., Taylor, T. E., Molyneux, M. E., Wirima, J. J., Vasalli,,I’., Hommel, M. & Lambert, P. H. (1989). Tumour necrosis factor and diseaseseverity in children with falciparum malaria. New England Journal of Medicine, 320, 1586 1591. Kwiatkowski, D., Hill, A. V. S., Sambou, I., Twumasi, I’., Castracane,J., Monogue, K. R., Cerami, A., Brewster, D. R. & Greenwood, B. M. (1990). TNF concentration in fatal cerebral, non-fatal cerebral and uncomplicated Plasmodium falciparum malaria. Lancer, 336, 1201-1204. Matsiota, I’., Balancher, A., Doyon, B., Guilbert, B., Clanet, M., Kouvelas, E. D. & Avrameas, S. (1988). Comparative study of natural autoantibodies in the serum and cerebrospinal fluid of normal individuals and patients with multiple sclerosisand other neurological disease.Annales de’1Znstitut Pasteurlhmunologie, 139,99-108. Mitra, S., Ravindran, B., Das, B. K., Das, R. K., Das, I’. K. & Rath, R. N. (1991). Human cerebral malaria: charac-
terization of malarial antibodies in cerebrospinalfluid. Clinical and Experimental Immunology,86, 19-21. Ramasamy, R. & Reese, R. T. (1986). Terminal galactose residues and the antigenicity of Plasmodium falciparum glycoproteins. Molecular and Biochemical Parasitology, 19, 91101. Ravindran, B., Satapathy, A. K. & Das, M. K. (1989). Naturally occurring anti-galactosyl antibodies in human Plasmodium falciparum infection-a possible role for autoantibodiesin malaria. Immunology Letters, 19, 137-142. Warrell, D. A., Looareesuwan, S., Phillips, R. E., White, N. J., Warrell, M. J., Chapel, H. M. & Tharavanij, S. (1986). Function of the blood-cerebrospinal fluid barrier in human cerebral malaria: rejection of the permeability hypothesis. AmericanJournal of Tropical Medicine and Hygiene, 35, 882889. Received 2 September 1991; accepted for publication 26 September 1991
Announcement
ROYAL
SOCIETY
OF TROPICAL
MEDICINE
AND
I-IYGIENE
The Transactions of the Royal Society of Tropical Medicine and Hygiene is issued bi-monthly (six parts per year) and publishes not only papers presented at Society meetings and symposia but also papers and correspondence submitted from all over the world on every aspect of tropical medicine and health. Supplements on specialist topics are published from time to time. The Transactions is available to Libraries and non-Fellows at an annual subscription of &96. It is dispatched by Accelerated Surface Post at no extra charge. The annual subscription to Fellows is &40. Applications for Fellowship must be made on the correct form, obtainable from the address below. Rate for bona fide students &25. Further
information
from: The Honorary Secretaries, Royal Society of Tropical Medicine and Hygiene, Manson House, 26 Portland Place, London WIN 4EY
