THE JOURNAL OF INFECTIOUS DISEASES • VOL. 134, NO.1· © 1976 by the University of Chicago. All rights reserved.
JULY 1976
Effect of Human Immune Serum Globulin on Infectivity of Hepatitis A Virus From the Department of Pediatrics, New York University School of Medicine, New York, New York
Saul Krugman
Table 1. Effect of immune serum globulin (lSG) on the infectivity of MS-l serum (type A) and MS-2 serum (type B).
In a previous report [1] we described the effect of standard human immune serum globulin (lSG) on the infectivity of MS-l serum containing hepatitis A virus and MS-2 serum containing hepatitis B virus. A standard ISG preparation that successfully neutralized the infecttivity of MS-l serum had no effect on the infectivity of MS-2 serum. The results of that study, which was conducted in 1969, are shown in table 1. The diagnosis of hepatitis in 1969 was based on biochemical evidence of liver dysfunction, such as abnormal serum transaminase values, either associated with or in the absence of jaundice. Since serologic tests for the detection of specific antibody were not available at that time, it was
Material inoculated MS-l (type A), no ISO MS-l (type A) plus ISO
No. of cases of primary hepatitis/no. of recipients 8/14* 0/10
NOTE. Table is modified from [1]. The ISO preparation used was Squibb lot no. 7-0. * The six recipients who did not have primary hepatitis A infection developed hepatitis after exposure as a result of contact with the first eight cases.
impossible (1) to determine the antibody content of the ISG preparation, (2) to confirm the presumed susceptibility of the participants in the study, or (3) to demonstrate the appearance of specific antibody. The recent development of a sensitive immune adherence HA (lAHA) test for the detection of hepatitis A antibody (anti-HA) [2] has enabled us to reevaluate the results of our earlier study. All serum specimens that had been stored in a deep freezer at - 20 C were tested under code for IAHA anti-HA by Dr. M. R. Hilleman of the Merck Institute for Therapeutic Research (West Point, Pa.).
Recieved for publication November 12, 1975, and in revised form January 12, 1976. This study was supported in part by contract no. DAMD 17-76-C-6012 from the U.S. Army Medical Research and Development Command. I am indebted to Dr. Maurice R. Hilleman and staff of the Merck Institute for Therapeutic Research for performing the immune adherence hemagglutination assays under code and to Harriet Friedman and Cass Lattimer for technical assistance. Please address requests for reprints to Dr. Saul Krugman, Department of Pediatrics, New York University Medical Center, 550 First Avenue, New York, New York 10016.
Materials and Methods
Protocol. The background of the study and the justification for carrying it out have been
70
Downloaded from http://jid.oxfordjournals.org/ at Freie Universitaet Berlin on June 28, 2015
A standard preparation of immune serum globulin containing a titer of immune adherence hemagglutination hepatitis A antibody of I: 3,200 neutralized the infectivity of MS-I serum. An inoculation of MS-I serum was followed by the following evidence of hepatitis A infection in eight of 14 seronegative recipients: (1) abnormal values for serum aspartate aminotransferase after an incubation period of 29-42 days, and (2) no detectable immune adherence hemagglutination hepatitis A antibody « 1:5) before exposure, and an eightfold or greater increase in antibody titer during convalescence. In contrast, inoculation of the mixture of MS-I serum and immune serum globulin was followed by (1) normal values for aspartate aminotransferase in all eight seronegative recipients, and (2) evidence of an antibody response (indicating subclinical infection) in two of the eight. Under the conditions of this study, the use of the preparation of immune serum globulin described prevented or modified hepatitis A infection.
ISG and Hepatitis A Virus Infectivity
Results
Titration of ISG. In the ISG preparation tested (lot no. 7-10; Squibb, New Brunswick, N. J.), the titer of anti-HA was 1:3,200, and the titer of anti-Hls, was 1: 8. Infectivity of MS-l serum. The serum specimens obtained before exposure from the 14 recipients of MS-1 serum had no detectable antiHA. As is indicated in table 2, evidence of type A hepatitis was detected in eight of 14 recipients 1 This dilution was erroneously reported to be 1: 10 in our earlier publication [1].
29-42 days after exposure. Biochemical evidence of liver dysfunction was associated with a fourfold or greater rise in IAHA anti-HA. Thus, an im inoculation of 0.1 ml of a 1: 2 dilution of MS-1 serum was infective for approximately 50% of the susceptible group. Transient jaundice was noted in only one of the eight children. The six susceptible children (no. 9-14, table 2) who were not infected by the MS-1 inoculum were exposed by contact with patients no. 1-8. All six children developed hepatitis ~27 days after contact exposure. Transient jaundice was noted in four of these six children. Neutralizing effect of ISG. The data in table 1 indicate that hepatitis was not detected in any of the 10 recipients of the MS-1 serumISG mixture. However, the serologic findings shown in table 3 revealed that two of the 10 children (no. 9A and lOB) had detectable antiHA prior to exposure. Consequently, eight of the 10 children were susceptible. All eight susceptible recipients of the MS-1 serum-ISG mixture were asymptomatic and had no evidence of liver dysfunction. However, two of the children (no. 7B and 8B) had serologic evidence of hepatitis A infection. Discussion
Under the conditions of this study, the use of a standard ISG preparation containing a titer of IAHA anti-HA of 1: 3,200 prevented or modified hepatitis A infection. All eight suspectible recipients of the MS-1 serum- ISG mixture were asymptomatic and had no biochemical evidence of liver dysfunction. Of this group two children had an inapparent infection, as indicated by the development of anti-HA in spite of normal results in tests of hepatic function (see below). These findings are very significant when they are compared with the effect of an inoculation of MS-1 serum without ISG. In these circumstances, the hepatitis that occurred in eight of 14 recipients was characterized by abnormal levels of SGOT as well as by an antibody response. This study highlighted the value of the IAHA antibody assay as a test for susceptibility or immunity. The two presumably susceptible children (no. 9A and lOB, table 3) had probably had unrecognized anicteric hepatitis A infection in the past.
Downloaded from http://jid.oxfordjournals.org/ at Freie Universitaet Berlin on June 28, 2015
described in detail [1]. Equal volumes of MS-1 serum (1: 2 dilution in distilled water) 1 and ISG were mixed and were incubated at 37 C for 1 hr and at 4 C for 1 hr. Ten presumably susceptible children received the mixture of MS-1 serum and ISG; the dose was 0.2 ml im. Fourteen presumably susceptible children received an im dose of 0.1 ml of MS-1 serum (1: 2 dilution) without ISG. Serial samples of serum were obtained before and for many weeks and months after the inoculation. The 24 children were studied as two separate groups, designated A and B. Group A included 14 children who were admitted to the unit in March 1969 and lived together for approximately three months after the study began on March 31, 1969. Of this group, nine children received MS-1 serum without ISG, and five received the MS-1 serum-ISG mixture. Group B included 10 children who were admitted in June 1969 and lived together for approximately three months after the study began on June 26, 1969. Children in group B had no contact with those in group A. Technical procedures. Serum bilirubin and serum aspartate aminotransferase (SGOT) tests of liver function were performed. A result was considered to be abnormal if the serum bilirubin value was > 1.0 mg/100 ml and if the SGOT value was ~60 units/ml. The details and significance of the test for IAHA anti-HA have been described previously [2, 3]. The passive HA (PHA) assay [4] was used to determine the content of antibody to hepatitis B surface antigen (anti-Hls.) in the ISG preparation.
71
Krugman
72
Table 2.
Clinical, biochemical, and serological findings in 14 children exposed to hepatitis A (MS-l strain). Peak serum bilirubin (mg/l00 ml)
Peak SGOT:j: (units/ml)
Interval from inoculation§
Anti-HA IAHA titerll
9
59
29
JULY 1976
Effect of Human Immune Serum Globulin on Infectivity of Hepatitis A Virus From the Department of Pediatrics, New York University School of Medicine, New York, New York
Saul Krugman
Table 1. Effect of immune serum globulin (lSG) on the infectivity of MS-l serum (type A) and MS-2 serum (type B).
In a previous report [1] we described the effect of standard human immune serum globulin (lSG) on the infectivity of MS-l serum containing hepatitis A virus and MS-2 serum containing hepatitis B virus. A standard ISG preparation that successfully neutralized the infecttivity of MS-l serum had no effect on the infectivity of MS-2 serum. The results of that study, which was conducted in 1969, are shown in table 1. The diagnosis of hepatitis in 1969 was based on biochemical evidence of liver dysfunction, such as abnormal serum transaminase values, either associated with or in the absence of jaundice. Since serologic tests for the detection of specific antibody were not available at that time, it was
Material inoculated MS-l (type A), no ISO MS-l (type A) plus ISO
No. of cases of primary hepatitis/no. of recipients 8/14* 0/10
NOTE. Table is modified from [1]. The ISO preparation used was Squibb lot no. 7-0. * The six recipients who did not have primary hepatitis A infection developed hepatitis after exposure as a result of contact with the first eight cases.
impossible (1) to determine the antibody content of the ISG preparation, (2) to confirm the presumed susceptibility of the participants in the study, or (3) to demonstrate the appearance of specific antibody. The recent development of a sensitive immune adherence HA (lAHA) test for the detection of hepatitis A antibody (anti-HA) [2] has enabled us to reevaluate the results of our earlier study. All serum specimens that had been stored in a deep freezer at - 20 C were tested under code for IAHA anti-HA by Dr. M. R. Hilleman of the Merck Institute for Therapeutic Research (West Point, Pa.).
Recieved for publication November 12, 1975, and in revised form January 12, 1976. This study was supported in part by contract no. DAMD 17-76-C-6012 from the U.S. Army Medical Research and Development Command. I am indebted to Dr. Maurice R. Hilleman and staff of the Merck Institute for Therapeutic Research for performing the immune adherence hemagglutination assays under code and to Harriet Friedman and Cass Lattimer for technical assistance. Please address requests for reprints to Dr. Saul Krugman, Department of Pediatrics, New York University Medical Center, 550 First Avenue, New York, New York 10016.
Materials and Methods
Protocol. The background of the study and the justification for carrying it out have been
70
Downloaded from http://jid.oxfordjournals.org/ at Freie Universitaet Berlin on June 28, 2015
A standard preparation of immune serum globulin containing a titer of immune adherence hemagglutination hepatitis A antibody of I: 3,200 neutralized the infectivity of MS-I serum. An inoculation of MS-I serum was followed by the following evidence of hepatitis A infection in eight of 14 seronegative recipients: (1) abnormal values for serum aspartate aminotransferase after an incubation period of 29-42 days, and (2) no detectable immune adherence hemagglutination hepatitis A antibody « 1:5) before exposure, and an eightfold or greater increase in antibody titer during convalescence. In contrast, inoculation of the mixture of MS-I serum and immune serum globulin was followed by (1) normal values for aspartate aminotransferase in all eight seronegative recipients, and (2) evidence of an antibody response (indicating subclinical infection) in two of the eight. Under the conditions of this study, the use of the preparation of immune serum globulin described prevented or modified hepatitis A infection.
ISG and Hepatitis A Virus Infectivity
Results
Titration of ISG. In the ISG preparation tested (lot no. 7-10; Squibb, New Brunswick, N. J.), the titer of anti-HA was 1:3,200, and the titer of anti-Hls, was 1: 8. Infectivity of MS-l serum. The serum specimens obtained before exposure from the 14 recipients of MS-1 serum had no detectable antiHA. As is indicated in table 2, evidence of type A hepatitis was detected in eight of 14 recipients 1 This dilution was erroneously reported to be 1: 10 in our earlier publication [1].
29-42 days after exposure. Biochemical evidence of liver dysfunction was associated with a fourfold or greater rise in IAHA anti-HA. Thus, an im inoculation of 0.1 ml of a 1: 2 dilution of MS-1 serum was infective for approximately 50% of the susceptible group. Transient jaundice was noted in only one of the eight children. The six susceptible children (no. 9-14, table 2) who were not infected by the MS-1 inoculum were exposed by contact with patients no. 1-8. All six children developed hepatitis ~27 days after contact exposure. Transient jaundice was noted in four of these six children. Neutralizing effect of ISG. The data in table 1 indicate that hepatitis was not detected in any of the 10 recipients of the MS-1 serumISG mixture. However, the serologic findings shown in table 3 revealed that two of the 10 children (no. 9A and lOB) had detectable antiHA prior to exposure. Consequently, eight of the 10 children were susceptible. All eight susceptible recipients of the MS-1 serum-ISG mixture were asymptomatic and had no evidence of liver dysfunction. However, two of the children (no. 7B and 8B) had serologic evidence of hepatitis A infection. Discussion
Under the conditions of this study, the use of a standard ISG preparation containing a titer of IAHA anti-HA of 1: 3,200 prevented or modified hepatitis A infection. All eight suspectible recipients of the MS-1 serum- ISG mixture were asymptomatic and had no biochemical evidence of liver dysfunction. Of this group two children had an inapparent infection, as indicated by the development of anti-HA in spite of normal results in tests of hepatic function (see below). These findings are very significant when they are compared with the effect of an inoculation of MS-1 serum without ISG. In these circumstances, the hepatitis that occurred in eight of 14 recipients was characterized by abnormal levels of SGOT as well as by an antibody response. This study highlighted the value of the IAHA antibody assay as a test for susceptibility or immunity. The two presumably susceptible children (no. 9A and lOB, table 3) had probably had unrecognized anicteric hepatitis A infection in the past.
Downloaded from http://jid.oxfordjournals.org/ at Freie Universitaet Berlin on June 28, 2015
described in detail [1]. Equal volumes of MS-1 serum (1: 2 dilution in distilled water) 1 and ISG were mixed and were incubated at 37 C for 1 hr and at 4 C for 1 hr. Ten presumably susceptible children received the mixture of MS-1 serum and ISG; the dose was 0.2 ml im. Fourteen presumably susceptible children received an im dose of 0.1 ml of MS-1 serum (1: 2 dilution) without ISG. Serial samples of serum were obtained before and for many weeks and months after the inoculation. The 24 children were studied as two separate groups, designated A and B. Group A included 14 children who were admitted to the unit in March 1969 and lived together for approximately three months after the study began on March 31, 1969. Of this group, nine children received MS-1 serum without ISG, and five received the MS-1 serum-ISG mixture. Group B included 10 children who were admitted in June 1969 and lived together for approximately three months after the study began on June 26, 1969. Children in group B had no contact with those in group A. Technical procedures. Serum bilirubin and serum aspartate aminotransferase (SGOT) tests of liver function were performed. A result was considered to be abnormal if the serum bilirubin value was > 1.0 mg/100 ml and if the SGOT value was ~60 units/ml. The details and significance of the test for IAHA anti-HA have been described previously [2, 3]. The passive HA (PHA) assay [4] was used to determine the content of antibody to hepatitis B surface antigen (anti-Hls.) in the ISG preparation.
71
Krugman
72
Table 2.
Clinical, biochemical, and serological findings in 14 children exposed to hepatitis A (MS-l strain). Peak serum bilirubin (mg/l00 ml)
Peak SGOT:j: (units/ml)
Interval from inoculation§
Anti-HA IAHA titerll
9
59
29
