Recent studies of human immunodeficiency virus type 1 (HIV-1) infection in humans and of simian immunodeficiency virus (SIV) in rhesus monkeys have shown that resolution of the acute viral infection and control of the subsequent persistent infection are mediated by the antiviral cellular immune response. We comparatively assessed several vaccine vector delivery systems-three formulations of a plasmid DNA vector, the modified vaccinia Ankara (MVA) virus, and a replication incompetent adenovirus type 5 (Ad5) vector-expressing the SIV gag protein for their ability to elicit such immune responses in monkeys. The vaccines were tested either as a single modality or in combined modality regimens. Here we show that the most effective responses were elicited by a replication-incompetent Ad5 vector, used either alone or as a booster inoculation after priming with a DNA vector. After challenge with a pathogenic HIV-SIV hybrid virus (SHIV), the animals immunized with Ad5 vector exhibited the most pronounced attenuation of the virus infection. The replication-defective adenovirus is a promising vaccine vector for development of an HIV-1 vaccine.

Human induced pluripotent stem (iPS) cells derived from somatic cells hold promise to develop novel patient-specific cell therapies and research models for inherited and acquired diseases. We and others previously reprogrammed human adherent cells, such as postnatal fibroblasts to iPS cells, which resemble adherent embryonic stem cells. Here we report derivation of iPS cells from postnatal human blood cells and the potential of these pluripotent cells for disease modeling. Multiple human iPS cell lines were generated from previously frozen cord blood or adult CD34 ؉ cells of healthy donors, and could be redirected to hematopoietic differentiation. Multiple iPS cell lines were also generated from peripheral blood CD34 ؉ cells of 2 patients with myeloproliferative disorders (MPDs) who acquired the JAK2-V617F somatic mutation in their blood cells. The MPD-derived iPS cells containing the mutation appeared normal in phenotypes, karyotype, and pluripotency. After directed hematopoietic differentiation, the MPD-iPS cell-derived hematopoietic progenitor (CD34 ؉CD45 IntroductionRecent derivation of human induced pluripotent stem (iPS) cells from patients' somatic cells has made it possible to generate patient-and disease-specific stem cell lines for developing novel cell therapies and disease modeling. 1-8 These human iPS cells exhibit characteristics similar to human embryonic stem (hES) cells, including unlimited expansion in culture. Using various vectors to deliver multiple transgenes encoding transcription factors, such as OCT4, SOX2, KLF4, and c-MYC, most published protocols were optimized to reprogram adherent cells, such as fibroblasts and keratinocytes from skin and hair. [1][2][3][4][5][6][7][8] It is also highly desirable to reprogram blood cells that are easily accessible and less exposed to environmental mutagens. For example, umbilical cord blood (CB) cells that are collected and stored in multiple cell banks could be used as a source of either autologous or allogeneic but histocompatible iPS cell lines. More critically, the ability to reprogram blood cells is essential if one wishes to generate iPS cells containing somatic mutations that are restricted to the blood cells and found only in acquired hematologic disorders to investigate their pathogenesis. A previous study demonstrated that differentiated mouse B cells could be reprogrammed to iPS cells, primarily using transgenic (reprogramming-ready) mice harboring the 4 reprogramming transgenes that are conditionally active. 9 More recently, mouse iPS cell lines were also derived from bone marrow (BM) progenitor cells obtained from a mouse whose hematopoiesis was reconstituted from a single congenic hematopoietic stem cell, providing further evidence that mouse hematopoietic cells can be reprogrammed to pluripotency. 10 Derivation of iPS cells from postnatal human blood cells has not been reported until recently when it was reported that granulocyte colony-stimulating factor (G-CSF) mobilized peripheral blood (PB) CD34 ϩ cells from a healthy person could...

Cellular immune responses, particularly those associated with CD3؉ CD8 ؉ cytotoxic T lymphocytes (CTL), play a primary role in controlling viral infection, including persistent infection with human immunodeficiency virus type 1 (HIV-1). Accordingly, recent HIV-1 vaccine research efforts have focused on establishing the optimal means of eliciting such antiviral CTL immune responses. We evaluated several DNA vaccine formulations, a modified vaccinia virus Ankara vector, and a replication-defective adenovirus serotype 5 (Ad5) vector, each expressing the same codon-optimized HIV-1 gag gene for immunogenicity in rhesus monkeys. The DNA vaccines were formulated with and without one of two chemical adjuvants (aluminum phosphate and CRL1005). The Ad5-gag vector was the most effective in eliciting anti-Gag CTL. The vaccine produced both CD4؉ and CD8 ؉ T-cell responses, with the latter consistently being the dominant component. To determine the effect of existing antiadenovirus immunity on Ad5-gag-induced immune responses, monkeys were exposed to adenovirus subtype 5 that did not encode antigen prior to immunization with Ad5-gag. The resulting anti-Gag T-cell responses were attenuated but not abolished. Regimens that involved priming with different DNA vaccine formulations followed by boosting with the adenovirus vector were also compared. Of the formulations tested, the DNA-CRL1005 vaccine primed T-cell responses most effectively and provided the best overall immune responses after boosting with Ad5-gag. These results are suggestive of an immunization strategy for humans that are centered on use of the adenovirus vector and in which existing adenovirus immunity may be overcome by combined immunization with adjuvanted DNA and adenovirus vector boosting.

Objective guidelines permitting safe return to sport following anterior cruciate ligament (ACL) reconstruction are infrequently used. The purpose of this study was to determine the published return to sport guidelines following ACL reconstruction in Level I randomized controlled trials. A systematic review was performed using Preferred Reporting Items for Systematic reviews and Meta-Analyses (PRISMA) guidelines. Level I randomized controlled trials were included if they reported a minimum 2-year follow-up after ACL reconstruction and return to sport criteria. Outcomes analyzed were the timing of initiation of return to sport, follow-up duration, and use of quantitative/qualitative criteria to determine return to sport. Forty-nine studies were included (N=4178; 68% male; mean patient age, 27.5±3.2 years; mean follow-up, 3.0±1.9 years; mean time from injury to reconstruction, 379±321 days). Ninety-six percent of reconstructions used autograft and 87% were single-bundle reconstructions. Lysholm score, single-leg hop, isokinetic strength, and KT-1000 or KT-2000 arthrometer (MEDmetric, San Diego, California) testing were performed in 67%, 31%, 31%, and 82% of studies, respectively. Only 5 studies reported whether patients were able to successfully return to sport. Ninety percent and 65% of studies failed to use objective criteria or any criteria, respectively, to permit return to sport. Description of permission/allowance to return to sport was highly variable and poor. Twenty-four percent of studies failed to report when patients were allowed return to sport without restrictions. Overall, 39%, 45%, and 51% of studies permitted running at 3 months, return to cutting/pivoting sports at 6 months, and return to sport without restrictions at 6 months, respectively. Further research into validated return to sport guidelines is necessary to fill the existing void in contemporary literature and to guide clinical practice.

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Septicemia is a frequent cause of death in HIV-infected adults in developing countries. Additional prospective studies are needed to determine the etiology of bloodstream infections (BSI) in febrile HIV-infected adults and guide initial evaluation and treatment in this setting. We assessed the prevalence and etiology of community-acquired BSI among 299 consecutive febrile adult medical admissions to Mulago Hospital, Kampala, Uganda, over a 4-month period in 1997. The median age of our patients was 30 years, 159 (53%) were male, and 227 (76%) HIV-1-seropositive. Overall, prevalence of bacteremia or fungemia (1 patient) was 24%. Bacteremia was more frequent in HIV-infected than in uninfected patients (27% versus 15%, respectively; p = .04). Mycobacterium tuberculosis (n = 28), Streptococcus pneumoniae (n = 15) and Salmonella species (n = 13) were the most frequent isolates. All Salmonella and mycobacterial isolates were recovered from HIV-infected patients. Pneumococcal bacteremia was not associated with HIV seropositivity. M. avium complex and M. simiae were isolated from two HIV-infected patients. The rate of mycobacteremia among febrile HIV-infected adults presenting for hospitalization was 13%. Bacteremia and disseminated tuberculosis are frequent causes of morbidity in febrile HIV-infected Ugandan adults. Initial empiric antibiotic coverage in this setting should be targeted toward the pneumococcus and gram-negative enteric bacilli, especially nontyphi Salmonella species. All patients presenting with chronic cough should be evaluated for tuberculosis.

The chronic myeloproliferative disorders (MPD) are clonal hematopoietic stem cell disorders of unknown etiology. We have reported defective thrombopoietin receptor (Mpl) protein expression in MPD patients. To determine whether the basis of abnormal Mpl protein expression was due to mutations in the Mpl gene, we sequenced Mpl cDNA from MPD patients. We found a single nucleotide substitution (G1238T) that results in a change from lysine to asparagine at amino acid 39 (K39N) in three AfricanAmerican women referred for an evaluation of an MPD. We subsequently screened more than 400 patients and controls and found that the K39N substitution is a polymorphism restricted to African Americans and that Ϸ7% of African Americans are heterozygous for K39N. African Americans with the K39N polymorphism had a significantly higher platelet count than controls without the polymorphism (P < 0.001) and reduced platelet protein Mpl expression. Expression of an Mpl cDNA containing the K39N substitution in cell lines was associated with incomplete processing and a reduction in Mpl protein, recapitulating the Mpl protein defect observed in platelets from individuals with K39N. K39N represents an identified functional Mpl polymorphism and is associated with altered protein expression of Mpl and a clinical phenotype of thrombocytosis.