424
LETTERS to the EDITOR
IVIG to prevent recurrent spontaneous abortion
Heat-shock proteins SIR,-Heat-shock proteins (HSP), whose synthesis is induced by increased body temperature, have moved from the esoteric to a role of clinical importance in areas as diverse as autoimmunity and cardiac protection (Lancet editorials of Sept 8 and Feb 2). The evolutionary conservation of these proteins from bacteria to man has raised the possibility that an immune response directed against the HSP of an invading microorganism will cross-react with the endogenous human protein, leading to autoimmunity. Bacterial HSP, such as those of mycobacteria, and antibodies directed against them have been widely used in research. Such reagents are very helpful in determining, for example, whether individual patients have antibodies or T cells directed against mycobacterial HSP and for looking at expression of the mycobacterial proteins in infected individuals. They thus allow important conclusions to be made about the role of bacterial infection in the aetiology of particular diseases. In contrast, however, extreme caution must be exercised in attempting to use these reagents to make conclusions about autoreactivity with the corresponding human HSP or to study the expression of the human proteins in uninfected individuals. The HSP of bacteria and man are closely related but they are not identical, and reagents directed against the bacterial protein will not necessarily react with the mammalian homologue. Much of the controversy in your columns about the role of the 65 kD HSP in insulin-dependent diabetes mellitus (Nov 17, p 1250; Jan 12, p 115) could have been avoided if Jones and colleagues1 had used a reagent directed against the human rather than the mycobacterial HSP. Thus the critical result in this report-that the binding of diabetes sera is inhibited by antibodies to mycobacterial 65 kD HSP-is now explained by the presence of mycobacterial antibodies in the sera. It has also been suggested that the antibody to the mycobacterial 65 kD protein may cross-react with human glutamic acid decarboxylase, which others have now identified as an autoantigen in diabetes? Both these difficulties would probably have been avoided by the use of an antibody specific to the human 65 kD HSP, whose expression and possible role in autoimmunity was being studied. Similarly, recent studies3 have indicated that T cells from rheumatoid arthritis patients that are able to react with mycobacterial HSP do not necessarily react with even the Escherichia coli homologue let alone the equivalent human protein. Therefore, although HSP are likely to play an important part in the aetiology of rheumatoid arthritis, the nature of the autoimmune reaction they provoke may be far more complex than first thought. Whilst studies using purified mycobacterial proteins and antibodies to them are of interest in examining the human immune response to these bacteria, extrapolation to the pathogenesis of human disease will require purified human HSP and specific antibodies. Fortunately, recombinant DNA techniques now permit the production of large amounts of purified protein, and monoclonal antibodies specific to human HSPs are widely available. The application of these reagents to human diseases as diverse as rheumatoid arthritis and myocardial infarction is an exciting prospect. Department of Biochemistry, Medical Molecular Biology Unit, UCMSM, University College London, London W1 P 6DP, UK
DAVID S. LATCHMAN
DB, Hunter NR, Duff GW Heat-shock protein 65 as a &bgr;-cell antigen of insulin-dependent diabetes Lancet 1990; 336: 583-85. 2. Backkeskov S, Aanstoot H-J, Chnstgau S, et al. Identification of the 64 k autoantigen m insulin-dependent diabetes as the GABA-synthesizing enzyme glutamic add decarboxylase. Nature 1990; 347: 151-56. 3. Winfield JB. Are heat shock proteins relevant to the pathogenesis of inflammatory arthritis? Clin Exp Rheumatol (in press) 1. Jones
SIR,-As an alternative to leucocyte immunotherapy, polyvalent intravenous immunoglobulin (IVIG) has been proposed12 to prevent recurrent spontaneous abortion (RSA).3 In a pilot study we showed that the success rate of IVIG was similar to that for leucocyte therapy.3 We present here an update on four years of experience with IVIG. 124 women with a history of primary (no living children, at least three first-trimester abortions) or secondary RSA (three or more miscarriages after delivery of healthy children) were investigated. Patients were considered for IVIG only if known causes of abortion such as chromosomal aberrations, gynaecological malformations, or hormonal disorders had been excluded. By November, 1990, 47 of these patients (35 primary, 12 secondary) became pregnant, and gave their informed consent for IVIG treatment. As soon as pregnancy was confirmed (usually by gestational week 5-6) the women were given 30 g IVIG (05-06 g/kg of ’Endobulin’ [Immuno]). Reflecting the half-life of IVIG, treatment was continued every 3 weeks with 20 g up to gestational week 25. RESULTS OF IVIG TREATMENT IN 47 PATIENTS WITH RSA
*Pregnancies without viable fetuses tCalculated from all completed pregnancies, includmg those still pregnant beyond week 25
Mild side-effects such as headache, slight fever, nausea, or were noted with 12 of the 223 IVIG infusions. The success rate was 75% for primary aborters and 60% for secondary ones (table). In primary RSA, IVIG appears to be as effective as leucocyte therapy, for which the reported success rates were 70-100%.1,2,4-6 In most unsuccessfully treated patients no sign of fetal viability could be detected by sonography. If these patients are excluded, on the assumption that no treatment can influence pregnancy outcome if there is no fetus, success rates would be 88% in primary and 86% in secondary cases. This implies that the protective effect of IVIG is equal in primary and in secondary RSA provided a viable fetus is present. Our results need confirmation by others and in larger numbers of patients. IVIG has several advantages over leucocyte therapy: the risk of viral infection or HLA immunisation is avoided; there is no need for treatment until pregnancy is confirmed, and IVIG can be used for patients with HLA antibodies. A randomised, placebocontrolled, double-blind multicentre trial of IVIG is now underway in Germany in patients with primary RSA.
tachycardia
Institute for Clinical Immunology and Transfusion Medicine, and Department of Obstetrics and Gynaecology, University of Giessen, 6300 Giessen, Germany
GERTRUD MUELLER-ECKHARDT OLAF HEINE BARBARA POLTEN
Taylor C, Faulk WP. Prevention of recurrent abortion with leucocyte transfusions Lancet 1981; ii: 68-70. 2. Beer AE, Quebbeman JF, Ayers JWZ, et al. Major histocompatibility complex antigens, maternal and paternal immune responses and chronic habitual abortions in humans. Am J Obstet Gynecol 1981; 141: 987-99. 3. Mueller-Eckhardt G, Heine O, Neppert J, et al. Prevention of recurrent spontaneous abortion by intravenous immunoglobulin. Vox Sang 1989, 56: 151-54. 1.
425
Mowbray JF, Gibbings C, Liddell H, et al. Controlled trial of treatment of recurrent spontaneous abortion by immunisation with paternal cells. Lancet 1985; i: 941-43. 5. Unander MA, Lindholm A, Olding LB. Blood transfusions generate/increase previously absent/weak blocking antibody in women with habitual abortion. Fertil Steril 1985, 44: 766-71. 6 Reznikoff-Etiévant MF, Duneux I, Huchet J, et al. Paternal leucocyte injections in recurrent spontaneous abortion. Lancet 1987; ii 1460. 4
New mutation in scrapie amyloid precursor gene (at codon 178) in Finnish Creutzfeldt-Jakob kindred SIR,-We have reported an association between a mutation :n codon 200 of the scrapie amyloid precursor gene (PRIP) and Creutzfeldt-Jakob disease (CJD) in patients of eastern European or Sephardic Jewish origin.1,2 In this latest (but certainly not last) chapter in the story of genetic abnormalities in CJD, we report a mutation in codon 178 linked to CJD in a large Finnish kindred. The pedigree now includes 15 affected members in four generations. The disease shows a trend towards "anticipation"; successive generations have had a progressively earlier age at onset,3 and the most recently affected member in the fourth generation is only 26 years old. Brain tissue from 1 patient transmitted disease to a capuchin monkey"; brain, spleen, and liver from another patient did not transmit disease to cats or other non-primates. DNA was extracted from frozen brain specimens of 2 affected family members, and the PRIP coding region was amplified by polymerase chain reaction and inserted into plasmid pGEM5z, from which multiple subclones were sequenced in both directions.S A G-to-A mutation in codon 178 (resulting in a substitution of asparagine for aspartic acid) was present in two of four subclones in 1 patient and all of five subclones in the other. This mutation was then screened by restriction endonuclease cleavage patterns in DNA extracted from frozen brain, spleen, anticoagulated blood, or blood clots from other affected and unaffected family members, unrelated sporadic CJD patients, and non-CJD controls. We used the enzyme TthlllI(New England Biolabs), visualising DNA fragment patterns by ethidium bromide after electrophoresis in 3% agarose gels. are shown in the figure. A single found in all 8 affected members (including both the transmitting and untransmitting cases), but not in any of 8 currently healthy first-degree relatives of the patients, 5 unrelated Finnish sporadic CJD patients, 12 Finnish controls (6 with other neurological disease, 6 with non-neurological illness), or 69 healthy North American controls. The clinical features of CJD in this family are in most respects typical of the familial disease described elsewhere. The mean age at
Representative patterns
mutation
was
is 50, and the illness evolves as a progressive dementia with associated cerebellar and upper motor neuron signs, muscular rigidity, and myoclonus. However, periodic EEG activity has not been observed, and the mean duration of illness of 21 months is longer than usual for either familial or sporadic CJD. onset
Neuropathological examination has revealed spongiform change, neuronal loss, and astroglial proliferation, without amyloid plaques. Thus, the codon 178 mutation in this family, like the codon 200 mutation, is clearly implicated in susceptibility to CJD, but does not seem to affect clinical course, pathological findings, or transmissibility. In contrast, the octapeptide repeat insertion found in two other CJD families is associated with unusual clinical heterogeneity,6 and codon 102 and 117 mutations are associated with the pathologically distinctive Gerstmann-StrausslerScheinker syndrome.7,8 As more patients with these mutations are studied, and additional mutations are found, a pattern may emerge that will
permit
a
correlation between molecular
genetics,
pathogenesis, and susceptibility in transmissible amyloidotic spongiform encephalopathies as a whole. Laboratory of CNS Studies and Laboratory of Molecular and Cellular Neurobiology (Section of Receptor Biochemistry and Molecular Biology), National Institute of Neurological Disorders and Stroke, National Institutes of Health,
Bethesda, Maryland 20892, USA, and Departments of Pathology and University of Helsinki,
Helsinki, Finland
LEV G. GOLDFARB MATTI HALTIA PAUL BROWN ANA NIETO JUSSI KOVANEN W. RICHARD MCCOMBIE Neurology,SUSAN TRAPP D. CARLETON GAJDUSEK
LG, Mitrová E, Brown P, et al. Mutation in codon 200 of scrapie amyloid protein gene in two clusters of Creutzfeldt-Jakob disease in Slovakia. Lancet 1990;
1. Goldfarb
336: 514. 2. Goldfarb LG, Korczyn AD, Brown P, et al. Mutation in codon 200 of scrapie amyloid precursor gene linked to Creutzfeldt-Jakob disease in Sephardic Jews of Libyan and non-Libyan origin. Lancet 1990; 336: 637. 3. Kovanen J, Haltia M. Descriptive epidemiology of Creutzfeldt-Jakob disease in Finland Acta Neurol Scand 1988; 77: 474-80. 4. Brown P, Rogers-Johnson P, Cathala F, et al. Creutzfeldt-Jakob disease of long duration: clinico-pathological characteristics, transmissibility, and differential diagnosis Ann Neurol 1984; 16: 295-304 5. Goldgaber D, Goldfarb LG, Brown P, et al. Mutations in familial Creutzfeldt-Jakob disease and Gerstmann-Straussler Scheinker syndrome. Exp Neurol 1989; 106: 204-06. 6. Owen F, Poulter M, Shah T, et al. An in-frame insertion in the prion protein gene in familial Creutzfeldt-Jakob disease. Mol Brain Res 1990; 7: 273-76. 7. Hsiao K, Baker HF, Crow TJ, et al. Linkage of a pnon missense variant to Gerstmann-Straussler-Scheinker syndrome. Nature 1989; 338: 342-45. 8. Tateishi J, Kitamoto T, Doh-ura K, et al. Immunochemical, molecular genetic, and transmission studies on a case of Gerstmann-Straussler-Scheinker syndrome. Neurology 1990; 40: 1578-81.
Seroepidemiological study of filovirus related to Ebola in the Philippines SIR,-Since November, 1989, several isolates of a filovirus closely related to Ebola virus have been recovered from cynomolgus monkeys (Macaca fascicularis) imported into the USA from the Philippines.1.2 The resemblance of the virus, tentatively
PRIP coding region fragments after
digestion
with Tth111 I.
CJD family S affected members (lanes 1-5);’ unaffected members (lanes 6-8), unrelated sporadic CJD cases (lanes 9, 10), non-CJD controls (lanes 11, 12) Tth111 1 cleavers the normal 803 bp PCR product into two fragments of 570 and 233 bp, the GAC-to-AAC mutation in codon 178 abolishes this cleavage site Individuals heterozygous for the mutation show these two fragments plus 803 bp (from uncleaved mutated allele)
named Reston strain, to Ebola virus is a concern because more than 600 registered cases with 71 % case fatality ratio caused two outbreaks in Sudan and Zaire in 1976.3>4 This has prompted the Philippine Department of Health to investigate the risk of transmission from animals to man and to assess the public health implications of this virus. We investigated all the export facilities and some wildlife trapping areas from December, 1989, to May, 1990. Most filovirus epizootics in a US quarantine facility were associated with monkeys from facility A in the Philippines. We therefore paid special attention to workers in the animal hospital at this facility, where the risk of exposure appeared highest. We did a serological survey of people exposed to cynomolgus monkeys in the Philippines. By interview we gathered information about possible risk factors for infection. Any history of recent illness, such as fever of sudden onset, diarrhoea, vomiting, and haemorrhagic manifestations, was also obtained. Sera were tested against Reston strain antigen in three laboratories (Research
LETTERS to the EDITOR
IVIG to prevent recurrent spontaneous abortion
Heat-shock proteins SIR,-Heat-shock proteins (HSP), whose synthesis is induced by increased body temperature, have moved from the esoteric to a role of clinical importance in areas as diverse as autoimmunity and cardiac protection (Lancet editorials of Sept 8 and Feb 2). The evolutionary conservation of these proteins from bacteria to man has raised the possibility that an immune response directed against the HSP of an invading microorganism will cross-react with the endogenous human protein, leading to autoimmunity. Bacterial HSP, such as those of mycobacteria, and antibodies directed against them have been widely used in research. Such reagents are very helpful in determining, for example, whether individual patients have antibodies or T cells directed against mycobacterial HSP and for looking at expression of the mycobacterial proteins in infected individuals. They thus allow important conclusions to be made about the role of bacterial infection in the aetiology of particular diseases. In contrast, however, extreme caution must be exercised in attempting to use these reagents to make conclusions about autoreactivity with the corresponding human HSP or to study the expression of the human proteins in uninfected individuals. The HSP of bacteria and man are closely related but they are not identical, and reagents directed against the bacterial protein will not necessarily react with the mammalian homologue. Much of the controversy in your columns about the role of the 65 kD HSP in insulin-dependent diabetes mellitus (Nov 17, p 1250; Jan 12, p 115) could have been avoided if Jones and colleagues1 had used a reagent directed against the human rather than the mycobacterial HSP. Thus the critical result in this report-that the binding of diabetes sera is inhibited by antibodies to mycobacterial 65 kD HSP-is now explained by the presence of mycobacterial antibodies in the sera. It has also been suggested that the antibody to the mycobacterial 65 kD protein may cross-react with human glutamic acid decarboxylase, which others have now identified as an autoantigen in diabetes? Both these difficulties would probably have been avoided by the use of an antibody specific to the human 65 kD HSP, whose expression and possible role in autoimmunity was being studied. Similarly, recent studies3 have indicated that T cells from rheumatoid arthritis patients that are able to react with mycobacterial HSP do not necessarily react with even the Escherichia coli homologue let alone the equivalent human protein. Therefore, although HSP are likely to play an important part in the aetiology of rheumatoid arthritis, the nature of the autoimmune reaction they provoke may be far more complex than first thought. Whilst studies using purified mycobacterial proteins and antibodies to them are of interest in examining the human immune response to these bacteria, extrapolation to the pathogenesis of human disease will require purified human HSP and specific antibodies. Fortunately, recombinant DNA techniques now permit the production of large amounts of purified protein, and monoclonal antibodies specific to human HSPs are widely available. The application of these reagents to human diseases as diverse as rheumatoid arthritis and myocardial infarction is an exciting prospect. Department of Biochemistry, Medical Molecular Biology Unit, UCMSM, University College London, London W1 P 6DP, UK
DAVID S. LATCHMAN
DB, Hunter NR, Duff GW Heat-shock protein 65 as a &bgr;-cell antigen of insulin-dependent diabetes Lancet 1990; 336: 583-85. 2. Backkeskov S, Aanstoot H-J, Chnstgau S, et al. Identification of the 64 k autoantigen m insulin-dependent diabetes as the GABA-synthesizing enzyme glutamic add decarboxylase. Nature 1990; 347: 151-56. 3. Winfield JB. Are heat shock proteins relevant to the pathogenesis of inflammatory arthritis? Clin Exp Rheumatol (in press) 1. Jones
SIR,-As an alternative to leucocyte immunotherapy, polyvalent intravenous immunoglobulin (IVIG) has been proposed12 to prevent recurrent spontaneous abortion (RSA).3 In a pilot study we showed that the success rate of IVIG was similar to that for leucocyte therapy.3 We present here an update on four years of experience with IVIG. 124 women with a history of primary (no living children, at least three first-trimester abortions) or secondary RSA (three or more miscarriages after delivery of healthy children) were investigated. Patients were considered for IVIG only if known causes of abortion such as chromosomal aberrations, gynaecological malformations, or hormonal disorders had been excluded. By November, 1990, 47 of these patients (35 primary, 12 secondary) became pregnant, and gave their informed consent for IVIG treatment. As soon as pregnancy was confirmed (usually by gestational week 5-6) the women were given 30 g IVIG (05-06 g/kg of ’Endobulin’ [Immuno]). Reflecting the half-life of IVIG, treatment was continued every 3 weeks with 20 g up to gestational week 25. RESULTS OF IVIG TREATMENT IN 47 PATIENTS WITH RSA
*Pregnancies without viable fetuses tCalculated from all completed pregnancies, includmg those still pregnant beyond week 25
Mild side-effects such as headache, slight fever, nausea, or were noted with 12 of the 223 IVIG infusions. The success rate was 75% for primary aborters and 60% for secondary ones (table). In primary RSA, IVIG appears to be as effective as leucocyte therapy, for which the reported success rates were 70-100%.1,2,4-6 In most unsuccessfully treated patients no sign of fetal viability could be detected by sonography. If these patients are excluded, on the assumption that no treatment can influence pregnancy outcome if there is no fetus, success rates would be 88% in primary and 86% in secondary cases. This implies that the protective effect of IVIG is equal in primary and in secondary RSA provided a viable fetus is present. Our results need confirmation by others and in larger numbers of patients. IVIG has several advantages over leucocyte therapy: the risk of viral infection or HLA immunisation is avoided; there is no need for treatment until pregnancy is confirmed, and IVIG can be used for patients with HLA antibodies. A randomised, placebocontrolled, double-blind multicentre trial of IVIG is now underway in Germany in patients with primary RSA.
tachycardia
Institute for Clinical Immunology and Transfusion Medicine, and Department of Obstetrics and Gynaecology, University of Giessen, 6300 Giessen, Germany
GERTRUD MUELLER-ECKHARDT OLAF HEINE BARBARA POLTEN
Taylor C, Faulk WP. Prevention of recurrent abortion with leucocyte transfusions Lancet 1981; ii: 68-70. 2. Beer AE, Quebbeman JF, Ayers JWZ, et al. Major histocompatibility complex antigens, maternal and paternal immune responses and chronic habitual abortions in humans. Am J Obstet Gynecol 1981; 141: 987-99. 3. Mueller-Eckhardt G, Heine O, Neppert J, et al. Prevention of recurrent spontaneous abortion by intravenous immunoglobulin. Vox Sang 1989, 56: 151-54. 1.
425
Mowbray JF, Gibbings C, Liddell H, et al. Controlled trial of treatment of recurrent spontaneous abortion by immunisation with paternal cells. Lancet 1985; i: 941-43. 5. Unander MA, Lindholm A, Olding LB. Blood transfusions generate/increase previously absent/weak blocking antibody in women with habitual abortion. Fertil Steril 1985, 44: 766-71. 6 Reznikoff-Etiévant MF, Duneux I, Huchet J, et al. Paternal leucocyte injections in recurrent spontaneous abortion. Lancet 1987; ii 1460. 4
New mutation in scrapie amyloid precursor gene (at codon 178) in Finnish Creutzfeldt-Jakob kindred SIR,-We have reported an association between a mutation :n codon 200 of the scrapie amyloid precursor gene (PRIP) and Creutzfeldt-Jakob disease (CJD) in patients of eastern European or Sephardic Jewish origin.1,2 In this latest (but certainly not last) chapter in the story of genetic abnormalities in CJD, we report a mutation in codon 178 linked to CJD in a large Finnish kindred. The pedigree now includes 15 affected members in four generations. The disease shows a trend towards "anticipation"; successive generations have had a progressively earlier age at onset,3 and the most recently affected member in the fourth generation is only 26 years old. Brain tissue from 1 patient transmitted disease to a capuchin monkey"; brain, spleen, and liver from another patient did not transmit disease to cats or other non-primates. DNA was extracted from frozen brain specimens of 2 affected family members, and the PRIP coding region was amplified by polymerase chain reaction and inserted into plasmid pGEM5z, from which multiple subclones were sequenced in both directions.S A G-to-A mutation in codon 178 (resulting in a substitution of asparagine for aspartic acid) was present in two of four subclones in 1 patient and all of five subclones in the other. This mutation was then screened by restriction endonuclease cleavage patterns in DNA extracted from frozen brain, spleen, anticoagulated blood, or blood clots from other affected and unaffected family members, unrelated sporadic CJD patients, and non-CJD controls. We used the enzyme TthlllI(New England Biolabs), visualising DNA fragment patterns by ethidium bromide after electrophoresis in 3% agarose gels. are shown in the figure. A single found in all 8 affected members (including both the transmitting and untransmitting cases), but not in any of 8 currently healthy first-degree relatives of the patients, 5 unrelated Finnish sporadic CJD patients, 12 Finnish controls (6 with other neurological disease, 6 with non-neurological illness), or 69 healthy North American controls. The clinical features of CJD in this family are in most respects typical of the familial disease described elsewhere. The mean age at
Representative patterns
mutation
was
is 50, and the illness evolves as a progressive dementia with associated cerebellar and upper motor neuron signs, muscular rigidity, and myoclonus. However, periodic EEG activity has not been observed, and the mean duration of illness of 21 months is longer than usual for either familial or sporadic CJD. onset
Neuropathological examination has revealed spongiform change, neuronal loss, and astroglial proliferation, without amyloid plaques. Thus, the codon 178 mutation in this family, like the codon 200 mutation, is clearly implicated in susceptibility to CJD, but does not seem to affect clinical course, pathological findings, or transmissibility. In contrast, the octapeptide repeat insertion found in two other CJD families is associated with unusual clinical heterogeneity,6 and codon 102 and 117 mutations are associated with the pathologically distinctive Gerstmann-StrausslerScheinker syndrome.7,8 As more patients with these mutations are studied, and additional mutations are found, a pattern may emerge that will
permit
a
correlation between molecular
genetics,
pathogenesis, and susceptibility in transmissible amyloidotic spongiform encephalopathies as a whole. Laboratory of CNS Studies and Laboratory of Molecular and Cellular Neurobiology (Section of Receptor Biochemistry and Molecular Biology), National Institute of Neurological Disorders and Stroke, National Institutes of Health,
Bethesda, Maryland 20892, USA, and Departments of Pathology and University of Helsinki,
Helsinki, Finland
LEV G. GOLDFARB MATTI HALTIA PAUL BROWN ANA NIETO JUSSI KOVANEN W. RICHARD MCCOMBIE Neurology,SUSAN TRAPP D. CARLETON GAJDUSEK
LG, Mitrová E, Brown P, et al. Mutation in codon 200 of scrapie amyloid protein gene in two clusters of Creutzfeldt-Jakob disease in Slovakia. Lancet 1990;
1. Goldfarb
336: 514. 2. Goldfarb LG, Korczyn AD, Brown P, et al. Mutation in codon 200 of scrapie amyloid precursor gene linked to Creutzfeldt-Jakob disease in Sephardic Jews of Libyan and non-Libyan origin. Lancet 1990; 336: 637. 3. Kovanen J, Haltia M. Descriptive epidemiology of Creutzfeldt-Jakob disease in Finland Acta Neurol Scand 1988; 77: 474-80. 4. Brown P, Rogers-Johnson P, Cathala F, et al. Creutzfeldt-Jakob disease of long duration: clinico-pathological characteristics, transmissibility, and differential diagnosis Ann Neurol 1984; 16: 295-304 5. Goldgaber D, Goldfarb LG, Brown P, et al. Mutations in familial Creutzfeldt-Jakob disease and Gerstmann-Straussler Scheinker syndrome. Exp Neurol 1989; 106: 204-06. 6. Owen F, Poulter M, Shah T, et al. An in-frame insertion in the prion protein gene in familial Creutzfeldt-Jakob disease. Mol Brain Res 1990; 7: 273-76. 7. Hsiao K, Baker HF, Crow TJ, et al. Linkage of a pnon missense variant to Gerstmann-Straussler-Scheinker syndrome. Nature 1989; 338: 342-45. 8. Tateishi J, Kitamoto T, Doh-ura K, et al. Immunochemical, molecular genetic, and transmission studies on a case of Gerstmann-Straussler-Scheinker syndrome. Neurology 1990; 40: 1578-81.
Seroepidemiological study of filovirus related to Ebola in the Philippines SIR,-Since November, 1989, several isolates of a filovirus closely related to Ebola virus have been recovered from cynomolgus monkeys (Macaca fascicularis) imported into the USA from the Philippines.1.2 The resemblance of the virus, tentatively
PRIP coding region fragments after
digestion
with Tth111 I.
CJD family S affected members (lanes 1-5);’ unaffected members (lanes 6-8), unrelated sporadic CJD cases (lanes 9, 10), non-CJD controls (lanes 11, 12) Tth111 1 cleavers the normal 803 bp PCR product into two fragments of 570 and 233 bp, the GAC-to-AAC mutation in codon 178 abolishes this cleavage site Individuals heterozygous for the mutation show these two fragments plus 803 bp (from uncleaved mutated allele)
named Reston strain, to Ebola virus is a concern because more than 600 registered cases with 71 % case fatality ratio caused two outbreaks in Sudan and Zaire in 1976.3>4 This has prompted the Philippine Department of Health to investigate the risk of transmission from animals to man and to assess the public health implications of this virus. We investigated all the export facilities and some wildlife trapping areas from December, 1989, to May, 1990. Most filovirus epizootics in a US quarantine facility were associated with monkeys from facility A in the Philippines. We therefore paid special attention to workers in the animal hospital at this facility, where the risk of exposure appeared highest. We did a serological survey of people exposed to cynomolgus monkeys in the Philippines. By interview we gathered information about possible risk factors for infection. Any history of recent illness, such as fever of sudden onset, diarrhoea, vomiting, and haemorrhagic manifestations, was also obtained. Sera were tested against Reston strain antigen in three laboratories (Research
