1056
ment-fixing (c.F.) antibody (titres 32, 64, and 128) and normal immunoglobulin concentrations. c.M.v. was cultured in human embryo lung fibroblasts, was detected by a cytopathic effect, and was identified by complement fixation. Urine samples-at least 4 taken before treatment-and others taken daily during treatment and weekly
Preliminary Communications TRANSFER-FACTOR TREATMENT IN CONGENITAL CYTOMEGALOVIRUS INFECTION I. T. THOMAS* G. T. HAWKINS
thereafter for 2-3 months were added to transport medium at 4°C and cultured within one hour. c.M.v. antibody was measured by complement fixation using AD 169 strain as antigen. Tuberculin P.P.D. 1/1000 (Weybridge) and candida antigen (1% Bencard DHS, 0.05 ml) were injected intradermally; nodules >10 mm for P.P.D. and >3 mm for candida 48 hours
J. F. SOOTHILL W. C. MARSHALL
Hospital for Sick Children and Departments of Immunology and Virology, Institute of Child Health, London When
Summary
two
doses of transfer factor
were
given to three infants with congenital cytomegalovirus (C.M.V.) infection, urine cultures for C.M.V. became negative, briefly, after five of the six doses.
days apart (see accompanying figure).
INTRODUCTION
ABOUT 0.3-1% of infants in developed countries are infected with cytomegalovirus (c.M.v.) in utero, and about 10% of these have gross brain and other damage;1,2 probably more have minor damage some of which may be progressive after birth.3.4 All excrete the virus in their urine for yearsS.6 and it would be desirable to stop the infection; however, idoxuridine, cytarabine, &c. have not succeeded in doing this.7 Recovery from virus infections probably depends on cell-mediated immunity (C.M.I.), which can be influenced by a dialysate of normal leucocytes (transfer factor [T.F.]).8The claim that T.F. was of benefit in intracellular bacterial, viral, and fungal infections9 led us to study the effect of T.F. on the excretion of c.M.v. by infants with congenital infection.
PATIENTS AND METHODS
based on symptoms and culture of c.M.v. from least 4 consecutive urine samples. We selected patients (aged 7-91 weeks) with evidence of definite brain damage, which was not severe enough to render useful improvement impossible. All showed delayed development, two had cerebral palsy, two had an occipitofrontal circumference smaller than the third percentile, one had seizures, and one had slight cerebral calcification. Other features included hepato-
Diagnosis
urine in
was
at
splenomegaly (2), thrombocytopenic purpura (2), neutropenia (1), and abnormal liver function (1). All had c.M.v. comple*Present address: Department of Pædiatrics, Merthyr Merthyr Tydfil, Glamorgan, Wales.
SKIN TEST AND LYMPHOCYTE RESPONSE
after injection were regarded as positive. Lymphocyte responses to phytohxmagglutinin (P.H.A.) (1, 2, 4, and 8 ug), tuberculin P.P.D. (Weybridge) (0-1, 1, 10, and 100 ug) and candida antigen (kindly supplied by Dr Mackenzie) (01, 1, 10, and 100 µg) were measured by thymidine uptake in 3 and 5 day cultures as described elsewhere.1O Results are expressed as stimulation index. T.F. was prepared" from blood of known blood-donors whose serum contained antibody to c.mt.v. One unit of T.F. (that from the leucocytes of 580 ml of blood) in 1 ml saline solution was injected subcutaneously in two dorsal sites, twice, 7-12
General
Hospital,
(PEAK STIMULATION
RESULTS
There were no adverse clinical effects after T.F. injections (except one episode of transient fever after the second dose of T.F.) or changes in blood or liver function tests. Both skin tests were negative in all three patients before T.F. (see accompanying table); patient 1 became positive to tuberculin and candida, and patient 3 to candida ; patient 2 remained skin-test-negative. Lymphocyte reactions to P.H.A. were normal in all three patients; patients 2 and 3 became positive to candida after T.F. (and patient 3 became sensitive to P.P.D.), but results of patient 1 were uninterpretable. c.M.v., which was cultured from all pre-treatment and late post-treatment urine samples, was not detected in one or more urine samples after all but one of the T.F. injections (see figure). Since the children were studied at different times, laboratory failure is unlikely to account for these negative cultures. The data are consistent with an effect starting 2 or more days after T.F. injection, and lasting up to 9 days, except for one late negative urine culture in patient 2. Negative urine culture was more common in the 3 weeks after the first injection (9 of 25) (p
ment-fixing (c.F.) antibody (titres 32, 64, and 128) and normal immunoglobulin concentrations. c.M.v. was cultured in human embryo lung fibroblasts, was detected by a cytopathic effect, and was identified by complement fixation. Urine samples-at least 4 taken before treatment-and others taken daily during treatment and weekly
Preliminary Communications TRANSFER-FACTOR TREATMENT IN CONGENITAL CYTOMEGALOVIRUS INFECTION I. T. THOMAS* G. T. HAWKINS
thereafter for 2-3 months were added to transport medium at 4°C and cultured within one hour. c.M.v. antibody was measured by complement fixation using AD 169 strain as antigen. Tuberculin P.P.D. 1/1000 (Weybridge) and candida antigen (1% Bencard DHS, 0.05 ml) were injected intradermally; nodules >10 mm for P.P.D. and >3 mm for candida 48 hours
J. F. SOOTHILL W. C. MARSHALL
Hospital for Sick Children and Departments of Immunology and Virology, Institute of Child Health, London When
Summary
two
doses of transfer factor
were
given to three infants with congenital cytomegalovirus (C.M.V.) infection, urine cultures for C.M.V. became negative, briefly, after five of the six doses.
days apart (see accompanying figure).
INTRODUCTION
ABOUT 0.3-1% of infants in developed countries are infected with cytomegalovirus (c.M.v.) in utero, and about 10% of these have gross brain and other damage;1,2 probably more have minor damage some of which may be progressive after birth.3.4 All excrete the virus in their urine for yearsS.6 and it would be desirable to stop the infection; however, idoxuridine, cytarabine, &c. have not succeeded in doing this.7 Recovery from virus infections probably depends on cell-mediated immunity (C.M.I.), which can be influenced by a dialysate of normal leucocytes (transfer factor [T.F.]).8The claim that T.F. was of benefit in intracellular bacterial, viral, and fungal infections9 led us to study the effect of T.F. on the excretion of c.M.v. by infants with congenital infection.
PATIENTS AND METHODS
based on symptoms and culture of c.M.v. from least 4 consecutive urine samples. We selected patients (aged 7-91 weeks) with evidence of definite brain damage, which was not severe enough to render useful improvement impossible. All showed delayed development, two had cerebral palsy, two had an occipitofrontal circumference smaller than the third percentile, one had seizures, and one had slight cerebral calcification. Other features included hepato-
Diagnosis
urine in
was
at
splenomegaly (2), thrombocytopenic purpura (2), neutropenia (1), and abnormal liver function (1). All had c.M.v. comple*Present address: Department of Pædiatrics, Merthyr Merthyr Tydfil, Glamorgan, Wales.
SKIN TEST AND LYMPHOCYTE RESPONSE
after injection were regarded as positive. Lymphocyte responses to phytohxmagglutinin (P.H.A.) (1, 2, 4, and 8 ug), tuberculin P.P.D. (Weybridge) (0-1, 1, 10, and 100 ug) and candida antigen (kindly supplied by Dr Mackenzie) (01, 1, 10, and 100 µg) were measured by thymidine uptake in 3 and 5 day cultures as described elsewhere.1O Results are expressed as stimulation index. T.F. was prepared" from blood of known blood-donors whose serum contained antibody to c.mt.v. One unit of T.F. (that from the leucocytes of 580 ml of blood) in 1 ml saline solution was injected subcutaneously in two dorsal sites, twice, 7-12
General
Hospital,
(PEAK STIMULATION
RESULTS
There were no adverse clinical effects after T.F. injections (except one episode of transient fever after the second dose of T.F.) or changes in blood or liver function tests. Both skin tests were negative in all three patients before T.F. (see accompanying table); patient 1 became positive to tuberculin and candida, and patient 3 to candida ; patient 2 remained skin-test-negative. Lymphocyte reactions to P.H.A. were normal in all three patients; patients 2 and 3 became positive to candida after T.F. (and patient 3 became sensitive to P.P.D.), but results of patient 1 were uninterpretable. c.M.v., which was cultured from all pre-treatment and late post-treatment urine samples, was not detected in one or more urine samples after all but one of the T.F. injections (see figure). Since the children were studied at different times, laboratory failure is unlikely to account for these negative cultures. The data are consistent with an effect starting 2 or more days after T.F. injection, and lasting up to 9 days, except for one late negative urine culture in patient 2. Negative urine culture was more common in the 3 weeks after the first injection (9 of 25) (p
