CD3+, CD56+ non-MHC restricted cytotoxic T lymphocytes in two fraternal AIDS patients: a case report.

We describe two brothers with hemophilia and AIDS, with an unusually large percentage of CD3+, CD56+ lymphocytes. They experienced no major complications with opportunistic infections (OI) and infrequent secondary infections, even though they had nearly 0% CD4 lymphocytes for 3 years. Both patients died in 1991 of progressive cardiomyopathy. The patients' lymphocytes were immunophenotyped by flow cytometry and analyzed for functional cytolytic activity against K562 and HIV infected HUT 78 cell lines. Single color CD4 counts were 2-9% for 4 years. Dual color CD4 counts at our laboratory demonstrated 0-1% CD4 for the last 6 months. When CD3+ lymphocytes were examined, both patient 1 and patient 2 demonstrated a significantly higher proportion and absolute number of CD3"bright"+, CD56+ double-positive cells, 47% and 22%, respectively, compared to other HIV-positive children with hemophilia (< or = 2%). Functional studies with the K562 target cell line demonstrated the highest natural killer (NK) lymphocyte activity in patient 1 that could not be augmented by in vitro addition of IL-2, whereas patient 2 had no NK activity unless IL-2 was added. Functional studies with HIV-infected HUT-78 cells demonstrated patient 2 had cytolytic activity against HIV-infected cells and patient 1 had high nonspecific cytolytic activity even against uninfected HUT-78 cells, whereas controls had minimal activity to HUT-78 cells or HIV-infected HUT-78 cells. The case report raises a speculative question requiring a larger database, whether the anti-HIV activity and/or unusual clinical course without typical O.I. of some AIDS patients may be related to the presence of CD3"bright"+, CD56+ lymphocytes of their immune system.

Analysis of a measles outbreak in Kent County, Michigan in 1990.

New recommendations from the Immunization Practices Advisory Committee call for a second dose of measles vaccine for school age children. The purpose of this paper is to demonstrate the need for additional measures for the protection of preschool age children. A hospital-based measles outbreak of 25 cases in Kent County, MI, in 1990 provided an opportunity to study measles transmission to preschool age children. Twenty-two (88%) were people who had never received measles vaccine. Twelve of the cases were unvaccinated preschoolers, seven of whom were older than 15 months. Three nonvaccinated, but eligible people (one philosophic exemption and two vaccine-eligible preschoolers) were the source of most of the other cases. One school age unvaccinated child died of measles pneumonitis. A telephone survey indicated that improved public education regarding indications and contraindications to vaccination might encourage vaccination of children according to public health recommendations.

HIV as a cause of giant cell hepatitis.

The 9-month-old daughter of human immunodeficiency virus (HIV)-seropositive parents presented with cholestasis and was found on liver biopsy to have giant cell hepatitis. No viral inclusions or particles were seen by light or electron microscopy. Ultrastructural studies of the liver biopsy demonstrated tubuloreticular structures in the endothelium and cylindrical confronting cisternae in inflammatory cells in the portal tracts. Serologic studies for hepatitis B, hepatitis A, and Epstein-Barr viruses were negative. Cytomegalovirus (CMV) was cultured from the urine, but buffy coat, nasopharyngeal, and liver cultures were negative and CMV antibody titer was low. The hepatitis responded dramatically to prednisone therapy. A repeat biopsy several months later revealed similar morphologic findings. AIDS was suspected on clinical and immunologic grounds, and was confirmed by the demonstration of HIV-specific IgG and IgM in serum. Five months after initial presentation, the infant developed Pneumocystis pneumonia, disseminated CMV infection, and died. This appears to be the first reported association of infantile giant cell hepatitis with HIV infection.

Children hospitalized with influenza B infection.

An outbreak of influenza virus type B infection occurred in Philadelphia from December, 1985, to April, 1986. During this epidemic 24 patients were admitted to Children's Hospital from whom influenza B was isolated from routine respiratory viral cultures. All were younger than 3 years of age. Clinical findings included fever (greater than or equal to 38 degrees C) (88%), rhinorrhea (62.6%), cough (50%), otitis (50%), rhonchi (42%), vomiting (38%), diarrhea (33%), rales (21%), pharyngitis (13%) and croup (4%). Remarkably 75% of the patients had underlying diseases which may have contributed to the severity of the infection. Nine (41%) patients had pneumonia. Two patients died of respiratory failure caused by overwhelming influenza B virus infection. Patients admitted to the hospital with respiratory and underlying diseases should have viral respiratory cultures which include influenza B.

Acquired immunodeficiency syndrome of childhood.

AIDS of childhood is reviewed in this timely article, including care of the child with infectious complications, and other current and future management concerns.

Toxic-shock syndrome associated with nasal packing: analogy to tampon-associated illness.

A pure culture of migmatoxin-producing Staphylococcus aureus was isolated from the purulent nasal packing removed from a 13-year-old premenarchal girl with toxic-shock syndrome after rhinoplasty. The similarities between this case and the tampon-associated illness are examined. An increased awareness of these similarities and the identification of predisposing factors may lead to earlier detection and treatment of toxic-shock syndrome.

Genetic analysis of transposon-induced mutations of the Rac prophage in Escherichia coli K-12 which affect expression and function of recE.

Fourteen mitomycin-resistant revertants of a recB21 recC22 strain were isolated after Tn5 mutagenesis. Eight of the mutations (type I) were essentially inseparable from aphA+ (Kanr) of Tn5; six (type II) were not. We hypothesize that the former are Tn5 and that the latter are IS50 insertions. Because of their phenotypic similarity to sbcA and sbcB mutations, which also suppress recB21 recC22, we have called them sbc mutations. sbc-lll::Tn5 was cotransducible with nirR and has thereby been located at position 29.8 on the Escherichia coli map in the vicinity of the Rac prophage and sbcA mutations. A recB21 recC22 sbc-lll::Tn5 strain was subjected to Tn10 mutagenesis, and a mitomycin- and UV-sensitive mutant was isolated. tet+ of Tn10 was 85% cotransducible with aphA+ of Tn5, locating these two transposons 0.1 map unit apart. A three-point cross located the Tn10 mutation at position 29.7. We hypothesize that the Tn10 insertion is located in recE and that the Tn5 and IS50 insertions activate expression of this gene. sbc-lll::Tn5 was found to be cis acting and dominant to its wild-type allele as were two sbcA mutations (sbcA1 and sbcA6). Five other type I and type II insertion mutations were dominant to their wild-type alleles. We hypothesize that the sbc insertion and sbcA mutations affect transcription regulation of recE and discuss the possibility that they do so differently.

Restriction nuclease and enzymatic analysis of transposon-induced mutations of the Rac prophage which affect expression and function of recE in Escherichia coli K-12.

Fourteen Tn5-generated mutations of the Rac prophage, called sbc because they suppress recB21 recC22, were found to fall into two distinct types: type I mutations, which were insertions of Tn5, and type II mutations, which were insertions of IS50. Both orientations of Tn5 and IS50 were represented among the mutants and were arbitrarily labeled A and B. All 14 of the Tn5 and IS50 insertions occurred in the same location (+/- 100 base pairs) approximately 5.6 kilobases from one of the hybrid attachment sites. Eleven of the mutants contained essentially the same amount of exonuclease VIII, the product of recE. The possibility that a promoter for recE was created by the insertion of Tn5 and IS50 was considered. Two IS50 mutants in which such a promoter could not have been created showed three to four times as much exonuclease VIII, and another showed one-half as much as the majority. The possibility was considered that a promoter internal to IS50 is responsible for this heterogeneity. Restriction alleviation was measured in all 14 mutants. An insertion of the transposon Tn10 which reduces expression of exonuclease VIII (recE101::Tn10) was located within the Rac prophage at a position 2.35 kilobases from the left hybrid attachment site. Location and orientation of the Rac prophage on the Escherichia coli genetic map are discussed in light of these results.

Insertion of transposons through the major cotransduction gap of Escherichia coli K-12.

The major cotransduction gap of the Escherichia coli chromosome extends from mini 31 to 34. We have inserted transposons through this gap which, by sequential transduction, link sbcA (min 29.8) with manA (min 35.7) and thus eliminate the gap. These results indicate that the length of DNA in the region, as measured by transduction, is not significantly different from the length obtained by conjugational time of entry. Since this segment of the E. coli chromosome has few known genes, these transposon insertions will be useful for genetic manipulations in the region of the gap. We describe the usefulness of these markers for rapidly mapping mutations which may be isolated in the region from min 27 to 37.

Transductional mapping of ksgB and a new Tn5-induced kasugamycin resistance gene, ksgD, in Escherichia coli K-12.

We have mapped the Escherichia coli ksgB gene to min 36.5, 0.8 min from man and 0.7 min from aroD. A new kasugamycin resistance (Ksgr) gene, ksgD, has been isolated, using a transposon, Tn5. ksgD::TN5 is 44% cotransducible with sbcA, unlinked to trp, and unlinked to man (by P1 transduction). The ksgD::Tn5 has a late time of entry from HfrB7 (PO43). These data place ksgD clockwise from sbcA (which enters early from HfrB7) at min 30.4. The reistance of ksgB ksgD single and double mutant strains has been quantitated. Single mutations, ksgB or ksgD, gave resistance to 600 micrograms of kasugamycin per ml, whereas a ksgB ksgD strain was able to grow in the presence of kasugamycin levels in excess of 3,000 micrograms/ml. This indicates that the mechanisms of resistance coded for by the two genes are independent and synergistic.

Repair of ultraviolet light-damaged deoxyribonucleic acid in sbc-A strains of Escherichia coli K-12.

An Escherichia coli strain carrying both rec+ and sbcA has been constructed. Repair of ultraviolet light-induced deoxyribonucleic acid damage was examined by measuring survival and thymine-dimer excision in the rec+ sbcA strain as well as rec+ sbcA+ and recB recC sbcA strains. The sbcA mutation restores normal survival in both recB recC uvrB and recB recC uvr+ strains. Excision of thymine-containing dimers does not occur in uvrB mutants, regardless of the rec or sbcA genotype. Survival, after ultraviolet-light damage, of a rec+ sbcA strain is quantitatively similar to rec+ sbcA+ and recB recC sbcA strains.

Metabolic characterization of the viable, residually dividing and nondividing cell classes of recombination-deficient strains of Escherichia coli.

The abilities of rec+, recB- recC-, recA-, and recA- recB- rec C- strains to support growth of bacteriophage T4, to take up oxygen, and to maintain cell integrity have been measured. (i) With respect to bacteriophage T4 growth, T4 phage is produced with identical lysis time in single -step growth curves with all strains tested. rec- strains show reduced phage production (fewer infected centers), but the average burst size per infected center is the same for all strains tested. Some rec- cells are unable to produce any phage, whereas the remainder of the rec-cells produce phage as rapidly and as efficiently as rec+ cells. (ii) With respect to oxygen consumption, rec- strains are deficient relative to the rec+ control to the same extent as the deficiency in phage production by theculture. The reduction in oxygen consumption is coordinate with reduction in rate of mass increase of the rec- culture. (iii) With respect to cell integrity, rec- cultures show increased lysis as measured by release of beta-galactosidase into the medium. The kinetics of release suggest that rec- nondividing cells all eventually lyse. These results are most consistent with the idea that rec- residually dividing cells and viable cells are metabolically normal according to the parameters we have measured, whereas nondividing cells are metabolically inactive.

The role of the rec genes in the viability of Escherichia coli K12.

Mutations in the recA, recB, or recC genes significantly reduce the growth rate and viability of Escherichia coli. Cultures of rec- strains are compose of three populations of cells: viable cells, nonviable but residually dividing cells, and nonviable and nondividing cells. Nondividing cells can be separated from dividing cells by penicillin treatment and velocity sedimentation. Nondividing cells of all rec- strains are greatly reduced in their ability to synthesize DNA. recB- recC- and recA- and recB- recC- nondividing cells contain DNA. This DNA synthesized in dividing cells and segregated into the nondividing cells. recA- nondividing cells contain little or no DNA. recA- recB- recC- nondividing cell DNA accumulates single-strand breaks.

Analysis of the growth of recombination-deficient strains of Escherichia coli K-12.

Cultures of recombination-deficient strains of Escherichia coli are composed of three classes of cells: (i) viable cells, which can undergo 20 or more generations, (ii) residually dividing cells, which can undergo fewer than 20 generations (probably an average of fewer than 6), and (iii) nondividing cells, which are incapable of a single division. The nonviable but residually dividing cells contribute to the mass increase of the culture, but not to the viability, thus accounting for the apparent dissimilarity in the effects of rec mutations on growth rates and viabilities. We have determined the frequencies of cells in each of the three classes, and, by making a simplifying assumption concerning the relative division times of viable and residually dividing cells, we have been able to describe mathematically the growth of the rec(-) cultures.