Klin. Pädialr. 203 (1991)
273
Seroprevalence and significance of antibodies to hepatitis C virus in pediatric patients with malignant diseases R. Schneppenheim l , P. Raulenberg 2 , H. Behnke l • 2, K. Vielor l 'Departments of Pediatrics and 2Microbiology, Christian-Albrechts-University, Kiel, 2300 Kiel I
The prevalence of antibodies to hepatitis C virus recombinant antigen c100-3 was determined in 82 pediatric patients with malignant diseases who received blood transfusions as support during their chemotherapy. By prescreening, 12 of 82 patients were repeatedly positive. No positive correlation was established neither with the number of transfused blood units nor with surrogate markers for hepatitis NA NB like ALT or anti HBc. Additional testing by alternate test assays revealed discordant results in II of these patients. Possible explanations, like reactivity to test-immanent coexpressed antigens of Saccharomyces cerevisae are discussed. Prävalenz und Relevanz von Anti-Hepatitis-C-Antikörpern bei pädiatrischen hämatologisch/onkologischen Patienten Die Prävalenz von Antikörpern gegen das rekombinante Hepatitis-C-Virus-Antigen c100-3 wurde bei 82 pädiatrischen hämatologisch/onkologischen Patienten ermittelt, die im Rahmen ihrer Chemotherapie eine unterschiedliche Anzahl von Blut-Transfusionen erhielten. 12 der 82 Patienten waren wiederholt seropositiv. Eine Korrelation zwischen diesem Ergebnis und der Anzahl der transfundierten Einheiten oder Surrogat-Markern der Hepatitis-NANB wie ALT und antiHBc ließ sich nicht nachweisen. Zusätzliche Tests mit anderen Test-Systemen ergaben in II Fällen abweichende Resultate. Eine mögliche Erklärung wäre die Reaktivität der untersuchten Seren mit Antigenen von Saccharomyces cerevisae, dem Expressionsorganismus des rekombinanten HCV-Antigens.
Introduction A significant number of pediatric patients treated with chemotherapy for malignant diseases require supportive therapy with red blood cell or platelet transfusions. Thus they are considered at risk for HCV infection. Classical groups being at risk or suspected to be infected Klin. Pädiatr. 203 (1991) 273-275 © 1991 F. Enke Verlag Stuttgart
with HCV comprise patients with hepatitis NANB (2), haemophilia and von Willebrand syndrome (10, 14, 15), i.v. drug abusers (6, 13), transplant recipients and patients on chronic dialysis (6, 11, 13). Since the development of a recombinant HCV ELlSA, an unexpected very high prevalence of antibodies (5-93070) against HCV antigen in these risk groups was reported (2, 6, 10, 11, 13, 14, 15). In European populations of healthy blood donors anti-HCV prevalence is only 0.3 to 1.40J0 (7, 8, 13, 16). However multitransfused patients are at an increased risk for HCV infection. Since in immunocompromised patients infections like post transfusion hepatitis might be of considerable significance, we retrospectively studied prevalence of anti HCV antibodies, correlation to surrogate markers of hepatitis NANB, morbidity corresponding to HCV status and possible transmission routes.
Patients and methods Eightytwo pediatric patients with a history of malignant disease were recruited for this study. Only 15 of them were still under therapy, i.e. in most cases under maintenance therapy of ALL or ANLL. Included were only patients having received blood transfusions to a cutoff point six months previously, to avoid falsely negative results because of a long incubation time. In order to avoid falseIy positive results, blood sampies were taken at least eight weeks after the last blood transfusions or treatment with intravenous immunoglobulin, which might be a significant source of anti HCV antibodies (Rautenberg unpublished). Sera of 82 pediatric patients with underlying malignant disease were prescreened by the Ortho HCV antibody Elisa test system, and initially reactive sampies were repeated in duplicate. Positive sera were additionally tested by the Abbott HCV EIA and the Chiron Riba HCV test, and follow-up testing was carried out several weeks later. All assays were used and interpreted as instructed by the manufacturers. Alanine aminotransferase (ALT) (ALT optimized method, Boehringer) and anti HBc (corzyme, AbboU Diagnostics) were determined as surrogate markers for hepatitis NANB. Other parameters tested were HBsAg (enzygnost HBsAg micro, Behring Werke) and HBV DNA (Hep probe DNA detection kit, Gibco/ BRL).
Downloaded by: NYU. Copyrighted material.
Summary
R. Schneppen heim el al.
Klin. Pädialr. 203 (1991) Since the recombinant HCV antigen c1003 is expressed in Saccharomyces cerevisae and coexpressed antigens 01' this microorganism could be a possible source for false positive results, sera 01' our patients were also tested for antibodies to Saccharomyces cerevisae and to another yeast, Candida albicans by an inhouse dot blot assay using SDS cell Iysates. Titers were determined by serial dilution. Antibody titers to Candida albicans > 3000 and to Saccharomyces cerevisae > 1000 were regarded as positive.
median end range of transtused unlts 30
24 25 20
2-64
10
In order to track possible transmission routes for HCV infection we recruted 86 out 01' ISO blood donors for anti HCV testing, who donated blood for our hcv seropositive patients.
No significant correlation was detected between seropositivity and the number 01' transfused blood units (s. figure I). Furthermore, none 01' the 86 blood do-
Tab. 1 Characteristics of children with malignant diseases according to their anti HCV status lürtho HCV EIISA)
__ _ _ _ _ _ _ _ _a_n_t_i_H_C_V positive no. of patients male/female antiHBcpos' ALT 20-50 U/L' ALT > 50 UIL' HBsAg pos HBV DNA pos •
=
In
12 6/6
negative
70 42/28
Ibtotal 82 48/34
3
3
6
3 1
12 4
15 5
0
4
4
0
0
0
the absence 01 HBsAg, HBV DNA, anti HBs
n-lI
HCY-poslt1ve
o
Ery. conc.
_
HCY-negat1ve n - 57
PlI. conc.
lall conc.
Fig. 1 Comparlson of the median erythrocyte and platelet transfusions In 68 reclplents tested positive or negative, respectlvely, for anti HCV antibodies Apparent dilferences between both groups were not slgnlflcant at P
273
Seroprevalence and significance of antibodies to hepatitis C virus in pediatric patients with malignant diseases R. Schneppenheim l , P. Raulenberg 2 , H. Behnke l • 2, K. Vielor l 'Departments of Pediatrics and 2Microbiology, Christian-Albrechts-University, Kiel, 2300 Kiel I
The prevalence of antibodies to hepatitis C virus recombinant antigen c100-3 was determined in 82 pediatric patients with malignant diseases who received blood transfusions as support during their chemotherapy. By prescreening, 12 of 82 patients were repeatedly positive. No positive correlation was established neither with the number of transfused blood units nor with surrogate markers for hepatitis NA NB like ALT or anti HBc. Additional testing by alternate test assays revealed discordant results in II of these patients. Possible explanations, like reactivity to test-immanent coexpressed antigens of Saccharomyces cerevisae are discussed. Prävalenz und Relevanz von Anti-Hepatitis-C-Antikörpern bei pädiatrischen hämatologisch/onkologischen Patienten Die Prävalenz von Antikörpern gegen das rekombinante Hepatitis-C-Virus-Antigen c100-3 wurde bei 82 pädiatrischen hämatologisch/onkologischen Patienten ermittelt, die im Rahmen ihrer Chemotherapie eine unterschiedliche Anzahl von Blut-Transfusionen erhielten. 12 der 82 Patienten waren wiederholt seropositiv. Eine Korrelation zwischen diesem Ergebnis und der Anzahl der transfundierten Einheiten oder Surrogat-Markern der Hepatitis-NANB wie ALT und antiHBc ließ sich nicht nachweisen. Zusätzliche Tests mit anderen Test-Systemen ergaben in II Fällen abweichende Resultate. Eine mögliche Erklärung wäre die Reaktivität der untersuchten Seren mit Antigenen von Saccharomyces cerevisae, dem Expressionsorganismus des rekombinanten HCV-Antigens.
Introduction A significant number of pediatric patients treated with chemotherapy for malignant diseases require supportive therapy with red blood cell or platelet transfusions. Thus they are considered at risk for HCV infection. Classical groups being at risk or suspected to be infected Klin. Pädiatr. 203 (1991) 273-275 © 1991 F. Enke Verlag Stuttgart
with HCV comprise patients with hepatitis NANB (2), haemophilia and von Willebrand syndrome (10, 14, 15), i.v. drug abusers (6, 13), transplant recipients and patients on chronic dialysis (6, 11, 13). Since the development of a recombinant HCV ELlSA, an unexpected very high prevalence of antibodies (5-93070) against HCV antigen in these risk groups was reported (2, 6, 10, 11, 13, 14, 15). In European populations of healthy blood donors anti-HCV prevalence is only 0.3 to 1.40J0 (7, 8, 13, 16). However multitransfused patients are at an increased risk for HCV infection. Since in immunocompromised patients infections like post transfusion hepatitis might be of considerable significance, we retrospectively studied prevalence of anti HCV antibodies, correlation to surrogate markers of hepatitis NANB, morbidity corresponding to HCV status and possible transmission routes.
Patients and methods Eightytwo pediatric patients with a history of malignant disease were recruited for this study. Only 15 of them were still under therapy, i.e. in most cases under maintenance therapy of ALL or ANLL. Included were only patients having received blood transfusions to a cutoff point six months previously, to avoid falsely negative results because of a long incubation time. In order to avoid falseIy positive results, blood sampies were taken at least eight weeks after the last blood transfusions or treatment with intravenous immunoglobulin, which might be a significant source of anti HCV antibodies (Rautenberg unpublished). Sera of 82 pediatric patients with underlying malignant disease were prescreened by the Ortho HCV antibody Elisa test system, and initially reactive sampies were repeated in duplicate. Positive sera were additionally tested by the Abbott HCV EIA and the Chiron Riba HCV test, and follow-up testing was carried out several weeks later. All assays were used and interpreted as instructed by the manufacturers. Alanine aminotransferase (ALT) (ALT optimized method, Boehringer) and anti HBc (corzyme, AbboU Diagnostics) were determined as surrogate markers for hepatitis NANB. Other parameters tested were HBsAg (enzygnost HBsAg micro, Behring Werke) and HBV DNA (Hep probe DNA detection kit, Gibco/ BRL).
Downloaded by: NYU. Copyrighted material.
Summary
R. Schneppen heim el al.
Klin. Pädialr. 203 (1991) Since the recombinant HCV antigen c1003 is expressed in Saccharomyces cerevisae and coexpressed antigens 01' this microorganism could be a possible source for false positive results, sera 01' our patients were also tested for antibodies to Saccharomyces cerevisae and to another yeast, Candida albicans by an inhouse dot blot assay using SDS cell Iysates. Titers were determined by serial dilution. Antibody titers to Candida albicans > 3000 and to Saccharomyces cerevisae > 1000 were regarded as positive.
median end range of transtused unlts 30
24 25 20
2-64
10
In order to track possible transmission routes for HCV infection we recruted 86 out 01' ISO blood donors for anti HCV testing, who donated blood for our hcv seropositive patients.
No significant correlation was detected between seropositivity and the number 01' transfused blood units (s. figure I). Furthermore, none 01' the 86 blood do-
Tab. 1 Characteristics of children with malignant diseases according to their anti HCV status lürtho HCV EIISA)
__ _ _ _ _ _ _ _ _a_n_t_i_H_C_V positive no. of patients male/female antiHBcpos' ALT 20-50 U/L' ALT > 50 UIL' HBsAg pos HBV DNA pos •
=
In
12 6/6
negative
70 42/28
Ibtotal 82 48/34
3
3
6
3 1
12 4
15 5
0
4
4
0
0
0
the absence 01 HBsAg, HBV DNA, anti HBs
n-lI
HCY-poslt1ve
o
Ery. conc.
_
HCY-negat1ve n - 57
PlI. conc.
lall conc.
Fig. 1 Comparlson of the median erythrocyte and platelet transfusions In 68 reclplents tested positive or negative, respectlvely, for anti HCV antibodies Apparent dilferences between both groups were not slgnlflcant at P
