Search

The human immunodeficiency virus epidemic continues in sub-Saharan Africa, and particularly affects adolescent girls and women who have limited access to antiretroviral therapy. Here we report that the risk of vaginal simian immunodeficiency virus (SIV)mac251 acquisition is reduced by more than 90% using a combination of a vaccine comprising V1-deleted (V2 enhanced) SIV envelope immunogens with topical treatment of the zinc-finger inhibitor SAMT-247. Following 14 weekly intravaginal exposures to the highly pathogenic SIVmac251, 80% of a cohort of 20 macaques vaccinated and treated with SAMT-247 remained uninfected. In an arm of 18 vaccinated-only animals without microbicide, 40% of macaques remained uninfected. The combined SAMT-247/vaccine regimen was significantly more effective than vaccination alone. By analysing immune correlates of protection, we show that, by increasing zinc availability, SAMT-247 increases natural killer cytotoxicity and monocyte efferocytosis, and decreases T-cell activation to augment vaccine-induced protection.

Posttransplantation cyclophosphamide (PTCy), given on days +3 and +4, reduces graft-versus-host disease (GVHD) after allogeneic hematopoietic cell transplantation (HCT), but its immunologic underpinnings are not fully understood. In a T-cell–replete, major histocompatibility complex-haploidentical murine HCT model (B6C3F1→B6D2F1), we previously showed that PTCy rapidly induces suppressive mechanisms sufficient to prevent GVHD induction by non-PTCy–exposed donor splenocytes infused on day +5. Here, in PTCy-treated mice, we found that depleting Foxp3+ regulatory T cells (Tregs) in the initial graft but not the day +5 splenocytes did not worsen GVHD, yet depleting Tregs in both cellular compartments led to fatal GVHD induced by the day +5 splenocytes. Hence, Tregs were necessary to control GVHD induced by new donor cells, but PTCy’s impact on Tregs appeared to be indirect. Therefore, we hypothesized that myeloid-derived suppressor cells (MDSCs) play a complementary role. Functionally suppressive granulocytic and monocytic MDSCs were increased in percentages in PTCy-treated mice, and MDSC percentages were increased after administering PTCy to patients undergoing HLA-haploidentical HCT. PTCy increased colony-stimulating factors critical for MDSC development and rapidly promoted the generation of MDSCs from bone marrow precursors. MDSC reduction via anti-Gr1 treatment in murine HCT did not worsen histopathologic GVHD but resulted in decreased Tregs and inferior survival. The clinical implications of these findings, including the potential impact of expanded MDSCs after PTCy on engraftment and cytokine release syndrome, remain to be elucidated. Moreover, the indirect effect that PTCy has on Tregs, which in turn play a necessary role in GVHD prevention by initially transplanted or subsequently infused T cells, requires further investigation.

Relapse limits the therapeutic efficacy both of chimeric antigen receptor (CAR) T cells and allogeneic hematopoietic cell transplantation (allo-HCT). Patients may undergo these therapies sequentially to prevent or treat relapsed malignancy. However, direct integration of the 2 therapies has been avoided over concerns for potential induction of graft-versus-host disease (GVHD) by allogeneic CAR T cells. We have shown in murine T-cell-replete MHC-haploidentical allo-HCT that suppressive mechanisms induced immediately after posttransplant cyclophosphamide (PTCy), given on days +3/+4, prevent GVHD induction by alloreactive T cells infused as early as day +5. Therefore, we hypothesized that allogeneic CAR T cells given in a similarly integrated manner in our murine MHC-haploidentical allo-HCT model may safely exert antitumor effects. Indeed, allogeneic anti-CD19 CAR T cells given early after (day +5) PTCy or even prior to (day 0) PTCy cleared leukemia without exacerbating the cytokine release syndrome occurring from the MHC-haploidentical allo-HCT or interfering with PTCy-mediated GVHD prevention. Meanwhile, CAR T-cell treatment on day +9 or day +14 was safe but less effective, suggesting a limited therapeutic window. CAR T cells infused before PTCy were not eliminated, but surviving CAR T cells continued to proliferate highly and expand despite PTCy. In comparison with infusion on day +5, CAR T-cell infusion on day 0 demonstrated superior clinical efficacy associated with earlier CAR T-cell expansion, higher phenotypic CAR T-cell activation, less CD4+CD25+Foxp3+ CAR T-cell recovery, and transcriptional changes suggesting increased activation of CD4+ CAR T cells and more cytotoxic CD8+ CAR T cells. This study provides mechanistic insight into PTCy’s impact on graft-versus-tumor immunity and describes novel approaches to integrate CAR T cells and allo-HCT that may compensate for deficiencies of each individual approach.

Background Selumetinib shrank inoperable symptomatic plexiform neurofibromas (PN) in children with neurofibromatosis type 1 (NF1) and provided clinical benefit for many in our previously published phase 1/2 clinical trials (SPRINT, NCT01362803). At the data cutoff (DCO) of the prior publications, 65% of participants were still receiving treatment. This report presents up to 5 years of additional safety and efficacy data from these studies. Methods This manuscript includes data from the phase 1 and phase 2, stratum 1 study which included participants with clinically significant PN-related morbidity. Participants received continuous selumetinib dosing (1 cycle = 28 days). Safety and efficacy data through February 27, 2021 are included. PN response assessed by volumetric magnetic resonance imaging analysis: Confirmed partial response (cPR) ≥20% decrease from baseline on 2 consecutive evaluations. Phase 2 participants completed patient-reported outcome measures assessing tumor pain intensity (Numeric Rating Scale-11) and interference of pain in daily life (pain interference index). Results For the 74 children (median age 10.3 years; range 3–18.5) enrolled, overall cPR rate was 70% (52/74); median duration of treatment was 57.5 cycles (range 1–100). Responses were generally sustained with 59% (44) lasting ≥ 12 cycles. Tumor pain intensity (n = 19, P = .015) and pain interference (n = 18, P = .0059) showed durable improvement from baseline to 48 cycles. No new safety signals were identified; however, some developed known selumetinib-related adverse events (AEs) for the first time after several years of treatment. Conclusions With up to 5 years of additional selumetinib treatment, most children with NF1-related PN had durable tumor shrinkage and sustained improvement in pain beyond that previously reported at 1 year. No new safety signals were identified; however, ongoing monitoring for known selumetinib-related AEs is needed while treatment continues.

S3. t-SLCO1B3 expression was optimized using various concentrations of chetomin at different timepoints in (A) LNCaP and (B) 22Rv1 cell lines.

Supplemental Figure Legends 1-4

Supplemental Table 1

Supplementary Figure 3 - PDF file 62K, Changes in mean heart rate during cycles 1 when patients taking betablockers were excluded

Supplementary Figure 4 - PDF file 314K, Changes in mean heart rate during cycles 1 when patients taking betablockers were excluded

Supplementary Figure 6 - PDF file 60K, Changes in heart rate observed for each patient in the NIH cohort during cycle 2 days 1, 8 and 15, excluding the patients taking beta-blockers

Supplementary Figure Legends - PDF file 25K, Legends describing supplementary figures

Supplemental Tables - PDF file 76K, Table S1. Diplotype arrangements of the ABCB1 SNPs calculated using Haploview. (Reproduced with authors' permission, Sissung et al, 2011(13); Table S2. Analysis of reported cardiovascular events for all 131 patients in NCI 1312; Table S3. Statistical Analyzes of Albumin, Potassium & Magnesium Data; Table S4. Analysis of variance to examine the relationship between albumin, potassium and magnesium levels in patients treated on NCI 1312; table legends

Supplementary Figure 5 - PDF file 60K, Changes in heart rate observed for patients in the NIH cohort during cycle 2 days 1, 8 and 15

Supplementary Figure 2 - PDF file 260K, Changes in mean heart rate occurring during cycle 1 days 1, 8 and 15 grouped by baseline heart rate

Supplementary Figure 7 - PDF file 317K, Changes in mean heart rate occurring during cycle 1 days 1, 8 and 15 grouped by baseline heart rate

Supplementary Figure 1 - PDF file 69K, Changes in heart rate observed for patients in the NIH cohort during cycle 1 days 1, 8 and 15

Supplementary Figure 8 - PDF file 299K, Changes in mean heart rate during cycles 2 when patients taking betablockers were excluded

Context The skeletal phenotype of patients with MEN2B has been described but fracture risk in these patients has not yet been evaluated. Objective This work aims to better delineate fracture risk in patients with multiple endocrine neoplasia type 2B (MEN2B). Methods This case series with chart review was conducted at the National Institutes of Health, Pediatric Oncology Branch. A total of 48 patients with MEN2B were identified, with an age range of 5 to 36 years, median of 19; 24 of 48 (50%) patients were female. Medical records, demographic information, available imaging, and laboratory results were reviewed. History up to age 19 was included in the statistical analyses. Results Of the 48 patients with MEN2B, 20 patients experienced at least one fracture. The majority (n = 18) experienced their first fracture at or before age 19. The observed frequency of fracture occurrence throughout childhood (0-19 years) was 38%, with very little difference between males and females. This frequency is higher than the 9.47 to 36.1 fractures per 1000 persons per year that has been reported in healthy pediatric cohorts in the United States. Less common sites of fracture including vertebral compression fracture and pelvic fractures were observed in patients with MEN2B. Conclusion In this group of patients with MEN2B, there was an increased overall risk of fracture compared to general pediatric cohorts in the United States. Less common sites of fracture were also observed. This suggests a possible effect of an activating RET mutation on bone physiology and warrants further investigation.

As demonstrated by the recent COVID pandemic, vaccines can reduce the burden arising from infectious agents. Adenoviruses (Ads) with deletion of the early region 1B55K (ΔE1B Ad) are currently being explored for use in vaccine delivery. Δ

Post-transplantation cyclophosphamide (PTCy) reduces the incidence and severity of graft-versus-host disease (GVHD), thereby improving the safety and accessibility of allogeneic hematopoietic cell transplantation (HCT). We have shown that PTCy works by inducing functional impairment and suppression of alloreactive T cells. We also have identified that reduced proliferation of alloreactive CD4

We investigated the impact of monocytes, NK cells, and CD8+ T-cells in primary HTLV-1 infection by depleting cell subsets and exposing macaques to either HTLV-1 wild type (HTLV-1WT) or to the HTLV-1p12KO mutant unable to infect replete animals due to a single point mutation in orf-I that inhibits its expression. The orf-I encoded p8/p12 proteins counteract cytotoxic NK and CD8+ T-cells and favor viral DNA persistence in monocytes. Double NK and CD8+ T-cells or CD8 depletion alone accelerated seroconversion in all animals exposed to HTLV-1WT. In contrast, HTLV-1p12KO infectivity was fully restored only when NK cells were also depleted, demonstrating a critical role of NK cells in primary infection. Monocyte/macrophage depletion resulted in accelerated seroconversion in all animals exposed to HTLV-1WT, but antibody titers to the virus were low and not sustained. Seroconversion did not occur in most animals exposed to HTLV-1p12KO. In vitro experiments in human primary monocytes or THP-1 cells comparing HTLV-1WT and HTLV-1p12KO demonstrated that orf-I expression is associated with inhibition of inflammasome activation in primary cells, with increased CD47 “don’t-eat-me” signal surface expression in virus infected cells and decreased monocyte engulfment of infected cells. Collectively, our data demonstrate a critical role for innate NK cells in primary infection and suggest a dual role of monocytes in primary infection. On one hand, orf-I expression increases the chances of viral transmission by sparing infected cells from efferocytosis, and on the other may protect the engulfed infected cells by modulating inflammasome activation. These data also suggest that, once infection is established, the stoichiometry of orf-I expression may contribute to the chronic inflammation observed in HTLV-1 infection by modulating monocyte efferocytosis.

Follicular CD8(+) T cells (fCD8 cells) reside within B cell follicles, believed to be immune privileged sites of HIV/SIV infection. We have observed comparable levels of fCD8 cells between chronically SIV infected rhesus macaques with low viral loads (LVL) and high viral loads (HVL), raising the question concerning their contribution to viremia control. Here we aimed to clarify the role of SIV-specific fCD8 cells in lymph nodes (LNs) over the course of SIV infection in rhesus macaques. We observed that fCD8, T follicular helper (Tfh), and T follicular regulatory (Tfreg) cells were all elevated in chronic SIV infection. fCD8 cells of LVL animals tended to express more Gag-specific granzyme B and exhibited significantly greater killing than HVL animals, and their cell frequencies negatively correlated with viremia, suggesting a role in viremia control. Env- and Gag-specific IL-21(+)Tfh of LVL but not HVL macaques negatively correlated with viral load, suggesting better provision of T cell help to fCD8 cells. Tfreg positively correlated with fCD8 cells in LVL animals and negatively correlated with viremia, suggesting a potential benefit of Tfregs via suppression of chronic inflammation. In contrast, in HVL macaques, Tfreg and fCD8 cell frequencies tended to negatively correlate, and a positive correlation was seen between Tfreg cell number and viremia, suggesting possible dysfunction and suppression of an effective fCD8 cell immune response. Our data suggest that control of virus-infected cells in B cell follicles not only depends on fCD8 cell cytotoxicity but also on complex fCD8 cell associations with Tfh and Tfreg.

To investigate the association between a cytochrome P450 17alpha-hydroxylase gene (CYP17) polymorphism and survival in Caucasian patients with androgen-independent prostate cancer (AIPC).The study used 222 samples acquired from Caucasian patients with AIPC. The CYP17 polymorphism (-34TC) was analyzed using polymerase chain reaction amplification followed by restriction fragment length polymorphism detection.No significant differences were observed in the frequencies of the CYP17 genotype in relation to categorized Gleason scores, age at diagnosis, or hormone therapy. The median survival was significantly longer in 126 patients with the CYP17 A2 allele (8.9 years) genotype than in 96 patients with the A1 allele (6.7 years) genotype (P = 0.040 by log-rank test). Similarly, the estimated survival probability at 10 years (24% in A1 allele versus 43% in A2 allele) showed a statistically significant difference between the two groups (P = 0.002 by the permutation test).These results suggest that the CYP17 polymorphism is associated with overall survival in patients with AIPC.

Two classes of two-event models for carcinogenesis are proposed. These models differ from some other multistage theories of carcinogenesis in that mutations areassociated with cell divisions, and the target tissue is allowed to grow in size. The first class of models is entirely stochastic, whereas in the second class of models, both deterministic and stochastic elements are involved. The incidence curves for childhood and adult cancers generated by these models are exhibited. Tests of the models against data were not undertaken. However, with the proper choice of parameters, these models generate incidence curves with the qualitative features of the curves seen in many common human carcinomas. Moreover the model proposed here for embryonal tumors is not distinguishable from the two-mutation model of Knudson et al. [6, 10, 11] on the basis of available data.

We present a mathematical treatment of a two-mutation model for carcinogenesis with time-dependent parameters. This model has previously been shown to be consistent with epidemiologic and experimental data. An approximate hazard function used in previous papers is critically evaluated.

Vital statistics data on mortality from malignant melanoma from five countries are simultaneously fit by the method of cohort analysis. After the effects of geographic and temporal trends are removed, the proportion of deaths occurring in the younger age groups is found to be higher in females than in males. Relative increases in mortality rates by birth cohort are approximately equal in different regions and appear to be slowing down in recent years.

SUMMARY Several parametric families of relative risk functions have been proposed as models for matched case-control and survival data. Some advantages accrue to those in which rela-tive risks are invariant under arbitrary translations of the origin of the covariate space, such as in the reassignment of values to dichotomous factors. It is shown in this paper that the family proposed by Guerrero & Johnson, which includes the commonly-used exponential and linear relative risk functions, has the simplest form of this origin invariance property. A differential equation that general origin-invariant relative risk functions must satisfy is derived. The effect of covariate translations on the parameter space is also discussed.

Mathematical expressions are derived, under different dosing patterns, for the number and size of premalignant clones within the framework of a two-mutation model for carcinogenesis, which has previously been shown to be consistent with a large body of epidemiologic and experimental data.

We apply results for the detection of outliers in logistic regression to the analysis of matched pairs data and illustrate the techniques with data from a matched pairs study of the relationship between iron-binding proteins and mortality in the Solomon Islands. Although the study includes 90 matched pairs, the conclusions change substantially with deletion of one or two pairs. Thus, even with reasonably large data sets, single observations may have a great impact on the results of an analysis. The identification of influential observations should constitute part of the analysis of matched case-control data.

Differential geometric methods are used to construct approximate confidence regions for curved exponential families. The $\alpha$-connection geometries discussed by Amari (1982), and another geometry introduced here, the $c$ geometry, are exploited to construct confidence regions. Survival and case-control studies with general relative risk functions are interpreted in the context of curved exponential families, and an example illustrates the construction of confidence regions for matched case-control studies. Simulations indicate that the geometric procedures have good coverage and power properties.

A geometric approach is described for the detection of influential points in nonlinear regression. This work extends and gives geometric interpretation to recent results in logistic regression diagnostics. An application of this approach to matched case-control studies is discussed.

The relative merits of the likelihood ratio statistic, the Wald statistic, and the score statistic are examined by an empirical evaluation based on matched case-control data. A mixture model for the relative-odds function is used. The likelihood ratio statistic is relatively constant for reasonable values of the mixture parameter, but the Wald statistic is unstable. The score statistic is shown to be independent of the mixture parameter. An exact expression is derived for the change in the score statistic upon deletion of risk sets, and an approximation is numerically evaluated.