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208 Abstracts Congenital Malformations at Michael Reese Hospital; Phatase, Peroxidase and Wright's Stain and Were Nor- 3

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208 Abstracts Congenital Malformations at Michael Reese Hospital; Phatase, Peroxidase and Wright's Stain and Were Nor- 3 208 Abstracts congenital malformations at Michael Reese Hospital; phatase, peroxidase and Wright's stain and were nor- 3. a similar clustering of chromosome abnormalities in mally vacuolated after bacterial phagocytosis. Normal newborns in Denver, Colorado (PUCK and ROBINSON, leukocyte motility and phagocytosis followed by de- Science [1965]) and in spontaneous abortions in Lon- granulation were seen with phase microscopy. De- don, Ontario (CARR, personal communication). For creased bactericidal activity of the patient's leukocytes the second cluster there was a corresponding increase during in vitro incubation with Staphylococcus aureus in the abortion population in Canada. There was no and Aerobacter aerogenes was repeatedly demonstrat- statistically significant increase in the reported incid- ed in the presence of the patient's serum or normal ence of 9 infectious diseases including rubella or infec- serum. To determine the nature of this defect a citric tious hepatitis, relative to the three clusterings defined acid extract of the patient's leukocyte granules was in the newborn population. Prospective studies on the compared with control specimens and decreased bac- distribution of chromosome aberration in time have tericidal activity was found. The deficient or abnormal been conducted since November, 1964, and reveals phagocytin activity of the patient's leukocytes, de- that when the spontaneous abortion and newborn po- monstrated by this study, could account for his clinical pulations are considered as one conceptual population, syndrome. (SPR) the distribution of chromosome aberrations in time appears random. These findings indicate that either 32 Defective Lymphocyte Response to PHA in Congenital the factors responsible for nonrandom distribution of Rubella. J.R. MONTGOMERY*, M.A. SOUTH*, chromosome aberrations were not operating during W.E.RAWLS*andJ.L.MELNiCK*,Depts.ofPed., the time period covered or that onrandom distribution Med., and Virology, Baylor Univ. College of of chromosome aberrations does not occur in the hu- Med.; and G.B.OLSEN*, P.B.DENT*, andR.A. man population. (APS) GOOD: Ped. Res. Labs., Variety Club Heart Hospital, U. of Minn., Minneapolis, Minn, 30 Chromosomes of Couples with Repeated Spontaneous (introduced by Martha D. Yow). Abortions. R. J. MCKAY, Jr., W.E. HODGKIN* and Persistent viral carrier state in congenital rubella E.H. WITTE*, University of Vermont College of remains an enigma. Defective cellular immune me- Medicine, Burlington, Vt. chanisms may play a role in this persistence. To Because a 20-25 % incidence of chromosomal ab- investigate this possibility the response of leukocytes normalities has been reported among spontaneously to phytohemagglutinin (PHA) was studied. 2xlO6 aborted fetuses, it was decided to study the chromo- peripheral leukocytes were cultured in routine media. somes of couples who have had 3 or more spontaneous PHA was added to achieve a concentration of 0.025 ml/ abortions. Using a commercial kit method for culturing ml of media. Quadruplicate cultures were incubated peripheral blood leukocytes, approximately 750-1000 at 37° C for 72 h and treated with C14 labeled thymidine metaphases were scanned in each patient in order to for 5 h prior to termination. Cellular response was obtain 50 satisfactory for counting. For each patient, estimated by measuring the cellular content of C14 in all metaphases with abnormal counts and 5 with nor- a standard liquid scintillation system. A decreased mal counts were photographed and karyotypes made. PHA responsiveness of leukocytes was demonstrated Among 4 couples with repeated abortions and one or in 8 of 14 congenital rubella patients studied, despite more children with multiple anomalies, 2 husbands evidence in these patients of normal delayed hyper- showed a balanced translocation in all metaphases sensitivity. PHA response returned to normal later in studied. Among the other 38 couples studied a number the course of each patient. To test whether this lack of showed abnormalities in a few cells (trisomy, partial responsiveness is due to an intrinsic defect in lymphoid trisomy, balanced and unbalanced translocations, cells from these patients or to direct effects of rubella fragments, dicentrics, endoreduplication, tetraploidy, virus, a normal adult's leukocytes were cultured with XO, long-armed Y, quadriradials and triradials). Par- rubella virus. PHA response was dramatically reduced tially trisomic cells were seen approximately 5 times as in every experiment; this effect could be eliminated by frequently among the women as among the men, and pretreatment of the virus preparation with rubella 6 patients were observed to have a single metaphase neutralizing antibody. Inhibition was also produced with a quadriradial chromosome. (APS) when Newcastle disease virus was substituted for ru- bella virus. These studies indicate that leukocytes from 31 The 'Impotent NeutrophiF Syndrome. L.L. KAHLE.* some congenital rubella patients show a defective re- H- MORENO*, and E. KAUDER*, Dept. of Pediat. sponse to PHA. The defective response can be repro- Univ. of Cincinnati, Cincinnati, O. (introduced duced in normal leukocytes by the addition of rubella by A.M. Mauer). virus or Newcastle disease virus in vitro. The prolonged Leukocyte function was studied in a 2-2,5-year-old persistence of virus and the defective reactivity to PHA Negro male with the history of repeated abscesses, cer- in babies with congenital rubella may be interrelated. vical and inguinal adenitis, recurrent pneumonia, se- (SPR) borrheic eczema of the scalp, hepatosplenomegaly and anemia. An appropriate leukocytosis with neutrophilia 33 A Mitotic Inhibitor Produced by Rubella Virus In- occurred with each infection, absolute neutrophil fection of Human Fibroblasts. STANLEY A. PLOTKIN, counts ranging from 3,900 to 24,500/mm3. IgG, IgA Wistar Institute and Department of Pediatrics, and IgM globulins were present in increased amounts. University of Pennsylvania, Philadelphia, Pa. Lymph node biopsy contained caseating granuloma. When infected with rubella virus, many human di- Skin tests and cultures for all types of acid fast bacilli ploid fibroblast cell strains show mild to marked de- and fungi were repeatedly negative. These findings are grees of mitotic inhibition (PLOTKIN et al.: Amer.J. characteristic of the syndrome of chronic granulomat- Epidem. 81: 71 [1965]); cell strains derived from in- ous disease of childhood. There was a normal flux of fected fetuses divide more slowly than normal (RAWLS leukocytes into Rebuck skin windows and exudate and MELNICK: J.exp.Med. 123: 795 [1966]); and in- fluid. Leukocytes appeared normal with alkaline phos- fants congenitally infected with rubella show mitotic Abstracts 209 inhibition and growth retardation (NAEYE and BLANC : cinees recorded daily temperatures and other symp- J.amer.med. Ass. 194: 1277 [1965]). In seeking an ex- toms on a card which was returned. Paired serum planation for these phenomena a substance inhibiting specimens were obtained at the time of vaccination mitosis was found in infected WI-38 cells. WI-38 cells and one month later from 1202 children. The results that had gone into mitotic arrest two weeks after ru- to date indicate: 1. 45 % of 592 children who had no bella virus infection were extracted by freeze-thawing. history of having mumps had detectable serum anti- When freed of virus, extracts of infected cells induced body before vaccination; 2. 98% of 340 seronegative mitotic inhibition of normal WI-38 cells and of a skin vaccinees had 4-fold or greater antibody response; fibroblast cell strain. No effect was seen on continuous 3. the vaccination was tolerated very well; the inci- lines of human cells or on non-human cell lines. The dence of febrile responses and of clinical symptoms was inhibitory substance was trypsin-sensitive and heat- essentially the same in 335 successfully vaccinated chil- labile but was unaffected by rubella antibody, ether, dren as compared with 252 children who were immune ribonuclease, or deoxyribonuclease. It was not sedi- prior to vaccination. These studies indicate that the mentable or acid-stable and did not protect cells against vaccine is antigenically potent and is clinically accep- the action of vesicular stomatitis virus. Concentrated table. (SPR) rubella virus inactivated by ultra-violet irradiation did not itself produce growth inhibition. 36 Experimental Genital Herpes Simplex infection in the This substance offers a potential explanation for the Mouse. ANDRE J.NAHMIAS, ZUHER M.NAIB*, mitotic inhibition associated with rubella virus infec- ANITA K. HIGHSMITH* and WILLIAM E.JOSEY*. tion in vitro and in vivo. (SPR) Emory Univ. Sch. of Med., Atlanta, Ga. In view of the important role of maternal genital 34 Clinical Experience with Natural and Atentuatedinfection as the source of herpes simplex virus (HSV) Rubella Virus Infection. H.M.MEYER, Jr.*, P.D. infection in the newborn (J.amer.med.Ass. 199: 1132 PARKMAN* and T.C.PANOS, National.Institutes [ 196 7]), an experimental model in mice was developed. of Health, Bethesda, Md., and Univ. of Arkan- Female mice could be readily infected by insertion in sas Medical Center, Little Rock, Ark. the vagina of a cotton pellet soaked with HSV. The Experience with attenuated rubella virus (strain occurrence of infection was substantiated by the re- HPV-77) indicates that infections in vaccinees are covery of virus from the vagina for as long as 12 days asymptomatic and non-communicable. HPV-77 is and by the demonstration
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