Post-treatment CD4 Research Articles

The efficacy of neoadjuvant immunotherapy and lymphocyte subset predictors in locally advanced esophageal squamous cell carcinoma: A retrospective study.

Despite the recognized therapeutic potential of programmed cell death protein 1/programmed death-ligand 1 (PD-1/PD-L1) inhibitors in advanced esophageal squamous cell carcinoma (ESCC), their role in neoadjuvant therapy and reliable efficacy biomarkers remain elusive. We retrospectively analyzed locally advanced ESCC patients who underwent surgery following a 2-cycle platinum and paclitaxel-based treatment, with or without PD-1 inhibitors (January 2020-March 2023). We assessed peripheral blood indexes and tertiary lymphoid structures (TLS) density to evaluate their impact on pathological response and prognosis, leading to a clinical prediction model for treatment efficacy and survival. Of the 157 patients recruited, 106 received immunochemotherapy (ICT) and 51 received chemotherapy (CT) alone. The ICT group demonstrated a superior pathological response rate (PRR) (47.2% vs. 29.4%, p = 0.034) with comparable adverse events and postoperative complications. The ICT group also showed a median disease-free survival (DFS) of 39.8 months, unattained by the CT group. The 1-year DFS and overall survival (OS) rates were 73% and 91% for the ICT group, and 68% and 81% for the CT group, respectively. We found higher baseline activated T cells, lower baseline Treg cells, and a decreased posttreatment total lymphocyte and CD4+/CD8+ ratio predicted an enhanced PRR. Reduced posttreatment CD4+/CD8+ ratio and increased NK cells were associated with prolonged survival, while higher TLS density indicated poorer prognosis. Among ICT group, a lower posttreatment CD4+/CD8+ ratio indicated longer DFS and reduced posttreatment B cells indicated longer OS. A nomogram integrating these predictors was developed to forecast treatment efficacy and survival. The combination of PD-1 inhibitors and chemotherapy appears promising for locally advanced ESCC. Evaluating the differentiation status and dynamic changes of peripheral blood immune cells may provide valuable predictive insights into treatment efficacy and prognosis.

Peripheral CD4+ T cells correlate with response and survival in patients with advanced non-small cell lung cancer receiving chemo-immunotherapy.

The aim of the present study was to explore the potential of peripheral immune cells in predicting the response and prognosis of patients with advanced non-small cell lung cancer (NSCLC) receiving anti-PD-1 immunotherapy and platinum-based chemotherapy. We utilized flow cytometry to examine the levels and dynamics of blood immune cells in 79 advanced NSCLC patients treated with the chemoimmunotherapy between December 2019 and January 2022. The pre- and post-treatment blood samples were collected within 3 days prior to the initiation of the first and third cycle of combination treatment, respectively. Progression-free survival (PFS) and overall survival (OS) analyses were conducted using Kaplan-Meier method and Cox regression models. The pre-treatment CD4+/Total T cells ratio was significantly higher in responders than non-responders (P < 0.05). The levels of pre-treatment total lymphocytes (P = 0.012), total B lymphocytes (P = 0.025), and NK cells (P = 0.022), and post-treatment NK cells (P = 0.011) and NKT cells (P = 0.035) were significantly associated with OS. Post-treatment CD8+/Total T cells ratio was positively correlated with OS (P = 0.038). In multivariate analysis, post-treatment NK cells and post-treatment CD4+CD8+/Total T cells ratio were negatively associated with OS (hazard ratio [HR] = 10.30, P = 0.038) and PFS (HR = 1.95, P = 0.022), respectively. Notably, significantly positive correlations were observed between CD4+/Total T cells ratio and prognosis both before and after treatment (P < 0.05). To summarize, our finding reveals that high CD4+/total T cells ratio was associated with favorable response and prognosis, highlighting its potential as a predictive biomarker to guide the selection of likely responders to platinum and anti-PD-1 combination therapy.

Development and Validation of a Three-Parameter Scoring System for Monitoring HIV/AIDS Patients in Low-Resource Settings Using Hematological Parameters.

This study aimed to test the validity of a composite score using complete blood count (CBC) for monitoring HIV patients receiving antiretroviral therapy (ART) in the absence of viral load and CD4 count. This retrospective cohort study analyzed the laboratory data of 82 HIV patients who had pre- and post-treatment viral load, CD4 count, and CBC data. Pre- and post-treatment data were pooled to analyze the correlation of CBC parameters with Polymerase Chain Reaction (PCR) ranks and their performance in indicating a CD4 count<200 cells/mm3 using the Operating Characteristics Curve (ROC), with the determination of cutoffs. A score combining the significant parameters was tested to predict a CD4 count of <200. Total lymphocyte count (TLC), percentage (TLP), and hemoglobin concentration (Hb) were the most significant parameters, showing negative correlations with PCR (Spearman's Rho = -0.357 to -0.242). The risk of acquired immunodeficiency syndrome (AIDS) was independently associated with TLC<1345 cells/mm3 (OR=2.92), TLP<29.07% (OR=3.53), and Hb<10.55 mg/dL (OR=3.60). A combined score of 2-3 indicated a CD4 count<200 with an odds ratio of 8.3-86.7. The proposed 3-parameter score combining the use of TLC, TLP, and Hb, is an affordable and practical approach that may have clinical utility in monitoring HIV patients receiving ART in low-resource settings.

Impact of Head and Neck Cancer Treatment on CD4 T Cell Decline and Recovery in People with HIV

The effect of cancer treatment on immunologic decline and recovery in people with HIV (PWH) is not well-described. This study aims to describe the trajectory of treatment-related immunosuppression in PWH treated for HNC predominantly with radiotherapy with or without chemotherapy and/or surgery. We performed a retrospective chart review of 23 PWH diagnosed with nonmetastatic HNC at our institution between 2002 and 2020. Baseline and longitudinal changes in CD4 count from the time of cancer treatment initiation were investigated. Patients were followed for 3 years after treatment completion. Nonparametric Mann-Whitney tests were used to compare median baseline CD4 count, post-treatment nadir CD4 count within 1 year after start of treatment, and peak recovery CD4 count within 3 years. Nonparametric Kruskal-Wallis and Mann-Whitney tests were used to compare median baseline, nadir, and peak recovery CD4 counts stratified by treatment. The majority of patients were male (20 [87%]), were white (13 [57%]), had a smoking history (21 [91%]), and had stage III-IV disease (17 [74%]). The median age was 58 (IQR 55-62). Primary subsites included oropharynx (11 [48%]), larynx (5 [22%]), oral cavity (4 [17%]), hypopharynx (2 [9%]) and nasal sinus (1 [4%]). Treatment comprised of surgery alone (S, n = 2), radiotherapy +/- surgery (RT, n = 6), and chemoradiotherapy +/- surgery (CRT, n = 15), thereby involving radiotherapy in 21 (91%) patients. All patients were on antiretroviral therapy at the time of diagnosis, and baseline HIV load was <200 copies/ml in 20 (87%) patients. Median baseline CD4 count prior to diagnosis was 592 cells/mm3 (IQR 329 to 834). The median nadir CD4 count was 201 cells/mm3 (IQR 104-282) or 34% of baseline (IQR 23-52%) (p<0.0001 compared to baseline), occurring at a median of 3.8 months (IQR 2.5-6.0). The median CD4 counts at 6 months and 1-year post-treatment were 236 cells/mm3 (IQR 135-337) and 274 cells/mm3 (IQR 150-334), respectively, representing 39% (IQR 31-58%) and 50% (IQR 40-63%) of baseline, respectively. The median peak recovery CD4 count was 431 cells/mm3 (IQR 371-561), or 68% of baseline (IQR 59-102%) (p<0.0001 compared to nadir), occurring at a median of 31 months (IQR 22-34). Stratified by treatment, the median nadir CD4 count as a percentage of baseline was 53% (S), 41% (RT), and 34% (CRT) (p = 0.32) and the peak recovery CD4 count as a percentage of baseline was 82% (S), 88% (RT), and 63% (CRT) (p = 0.45). Among PWH with HNC, CD4 count nadirs to a 1/3 of baseline but gradually rebounds to 2/3 of baseline over the subsequent 2-3 years. While this data supports continuing to treat HNC in PWH with standard-of-care management, CD4 count should be monitored closely for several years following HNC radiotherapy.

Gastrodin Improves Quality of Life and Outcome of Autoimmune Encephalitis Treatment with High-Dose Gamma Globulin and Collaborative Care

This study aims to elucidate the effectiveness of gastrodin in improving the outcome of autoimmune encephalitis patients receiving high-dose gamma globulin plus collaborative nursing care. Gastrodin led to higher post-treatment Mini-mental State Examination scores, CD3+, CD4+, IgA, IgG, and IgM than the control group, and lower inflammatory cytokine levels (P &lt; 0.05). To sum up, gastrodin administration to patients receiving high-dose gamma globulin and collaborative nursing care was highly effective in treating autoimmune encephalitis and can effectively improve patients’ quality of life.

Effects of Gender and Baseline CD4 Count on Post-Treatment CD4 Count Recovery and Outcomes in Patients with Advanced HIV Disease: A Retrospective Cohort Study.

Presentation to care with advanced HIV disease (AHD) is a significant problem in sub-Saharan Africa. We evaluated factors associated with immune recovery among individuals presenting to care with AHD in Zimbabwe. We conducted a retrospective evaluation of outcomes among adult (>18 years old) individuals with AHD (CD4 count ≤200 cells/mm3) receiving care at 18 outpatient primary care clinics in Harare, Zimbabwe. Baseline and 12-month CD4 count data were extracted from medical records. CD4 count recovery (defined as CD4 count >200 cells/mm3) after 12 months on non-nucleotide reverse transcriptase inhibitor (NNRTI)-based antiretroviral therapy (ART) regimen was determined and factors associated with CD4 count recovery were established using logistic regression. All statistical analysis was performed on SPSS v23. A total of 1,338 participant records were included in the analysis. The median interquartile range (IQR) age was 37 (30-43) years and 52% were females. The baseline median (IQR) CD4 count was 50 (28-75) cells/mm3 and was significantly lower among patients with history of cryptococcal meningitis compared to those without [25 (10-52) vs. 52 (32-77), respectively; p = .0009]. The median (IQR) CD4 count at 12 months after ART initiation increased from 50 (28-75) at baseline to 180 (92-290) cells/mm3. Immune recovery with a CD4 count >200 cells/mm3 was observed in 181/417 (43%). Male gender and low baseline CD4 count were strong predictors of poor immunological recovery on ART. Immunological recovery following ART initiation was 43% among individuals with AHD. Male patients are most vulnerable to persistent immunological failure. Clinical Trial Registration number: NCT02434172.

Intracellular IFN-γ and IL-4 levels of CD4 + and CD8 + T cells in the peripheral blood of naturally infected (Leishmania infantum) symptomatic dogs before and following a 4-week treatment with miltefosine and allopurinol: a double-blinded, controlled and cross-sectional study

BackgroundCanine leishmaniosis (CanL) is a systemic disease caused by the protozoan parasite Leishmania infantum with a wide spectrum of clinical signs, with cutaneous, ocular, renal and lymphoreactive conditions prevailing in the clinical setting. The immune system plays a pivotal role in the evolution of Leishmania infection and its response to antileishmanial treatment. Cytokines are important immune response mediators that are released by activated lymphocytes and less so by other immunocytes. In dogs with leishmaniosis, IFN-γ and IL-4 have been recognized as the main activators of cellular and humoral immunity, respectively. The objective of this study was to investigate intracellular IL-4 and IFN-γ expression by CD4 + and CD8 + lymphocytes in the peripheral blood of symptomatic dogs before and after combined antileishmanial treatment with miltefosine and allopurinol.ResultsPostantileishmanial treatment CD4 + IL-4 + and CD8 + IL-4 + cell counts were significantly decreased, although no similar changes were observed in the comparisons made between the pre- and posttreatment CD4 + IFN-γ + and CD8 + IFN-γ + counts and ratios.ConclusionThe findings indicate that IL-4 production by T cells may facilitate the symptomatic phase of CanL, whereas IFN-γ production by CD4 + and CD8 + cells may indicate its negligible role in the evolution of natural CanL and perhaps the equivocal positive influence of antileishmanial treatment.

442. Clinical Outcomes of Checkpoint Inhibitor Therapy in People with HIV: A Single-Center Case Series

Abstract Background Immunotherapeutics, particularly immune-checkpoint inhibitors (ICI), have transformed the treatment landscape for advanced cancer. Such therapies could considerably benefit people with HIV (PWH), yet PWH have often been excluded from ICI trials such that data regarding ICI safety in PWH are limited. In addition, ICIs have recently been proposed as latency reversing agents that could impact the HIV reservoir. Here, we report the outcomes in a cohort of PWH after ICI therapy. Methods Using the electronic health record, we systematically identified all PWH with advanced cancer who received ICIs from January 2000 to December 2018 at one academic medical center. We collected data on demographics, HIV history, and HIV and hematologic parameters before and after ICI in 25 PWH over 29 treatment episodes. We cataloged adverse events following treatment. Results In this cohort, ICI therapy was not associated with a significant increase in plasma HIV RNA levels nor with significant change in CD4+ T-cell count, CD8+ T-cell count, or CD4/CD8 ratio (Figure 1). Among 19 treatment episodes with available pre- and post- treatment data, there were five viral blips (26%); HIV RNA exceeded 200 copies/mL in one episode. The blip rate observed among PWH on ART in the literature ranges from 10-30% and among PWH receiving ICI from 0-33%. Adverse events included infectious and autoimmune conditions across multiple organ systems including systemic, pulmonary, gastrointestinal/hepatic, genitourinary, neurologic, endocrine, and dermatologic. The most common adverse events were diarrhea (n=7) and rash (n=5). Five deaths occurred within one month of ICI initiation; all were attributed to progressive malignancy. Figure 1:Changes in HIV parameters associated with checkpoint inhibitor therapy (a) Pretreatment plasma HIV RNA, n=23. There was one episode in which the pretreatment plasma HIV RNA was greater than 200 copies/ml. (b) Pretreatment CD4+ Tcell counts, n=24. (c) Longitudinal plasma HIV RNA trends in 5 individuals who experienced a viral blip, defined as an increase in plasma HIV RNA from “not detected” to “detected” or greater within a year of ICI initiation. Symbols represent unique participants. (d) Pretreatment and posttreatment plasma HIV RNA values closest to ICI initiation, n=19. Points are scaled to proportion of sample. (e) Pretreatment and posttreatment CD4+ T-cell counts closest to ICI initiation, n=20. Thick line represents the change in median M between the two time points. (f) Pretreatment and posttreatment CD4+ T-cell percentages closest to ICI initiation, n=19.Thick line represents the change in median M between the two time points. (g) Pretreatment and posttreatment CD8+ T-cell counts closest to ICI initiation, n=18. Thick line represents change in median M between the two time points. (h) Pretreatment and posttreatment CD4/CD8 ratios closest to ICI initiation, n=18. Thick line represents the change in median M between the two time points. Conclusion This single-center cohort demonstrated no significant impact on HIV parameters among PWH receiving ICI. This study contributes to the growing body of evidence suggesting the safety and tolerability of ICI in PWH in both HIV and non-HIV-related outcomes and supports the inclusion of PWH in future ICI clinical trials. While we did not see an increased rate of viral blips on routine clinical testing, our findings also provide key preliminary data supporting the safety and tolerability of ICIs in future studies of these agents as HIV therapeutics, for example as agents that may reverse HIV latency. Disclosures All Authors: No reported disclosures.

Baseline Characteristics of Mitochondrial DNA and Mutations Associated With Short-Term Posttreatment CD4+T-Cell Recovery in Chinese People With HIV.

BackgroundMitochondrial DNA (mtDNA) profiles and contributions of mtDNA variants to CD4+T-cell recovery in Euramerican people living with HIV (PLWH) may not be transferred to East-Asian PLWH, highlighting the need to consider more regional studies. We aimed to identify mtDNA characteristics and mutations that explain the variability of short-term CD4+T-cell recovery in East-Asian PLWH.MethodEight hundred fifty-six newly reported antiretroviral therapy (ART)-naïve Chinese PLWH from the Comparative HIV and Aging Research in Taizhou (CHART) cohort (Zhejiang Province, Eastern China) were enrolled. MtDNA was extracted from peripheral whole blood of those PLWH at HIV diagnosis, amplified, and sequenced using polymerase chain reaction and gene array. Characterization metrics such as mutational diversity and momentum were developed to delineate baseline mtDNA mutational patterns in ART-naïve PLWH. The associations between mtDNA genome-wide single nucleotide variants and CD4+T-cell recovery after short-term (within ~48 weeks) ART in 724 PLWH were examined using bootstrapping median regressions.ResultsOf 856 participants, 74.18% and 25.82% were male and female, respectively. The median age was 37 years; 94.51% were of the major Han ethnicity, and 69.04% and 28.62% were of the heterosexual and homosexual transmission, respectively. We identified 2,352 types of mtDNA mutations and mtDNA regions D-loop, ND5, CYB, or RNR1 with highest mutational diversity or volume. Female PLWH rather than male PLWH at the baseline showed remarkable age-related uptrends of momentum and mutational diversity as well as correlations between CD4+T <200 (cells/μl) and age-related uptrends of mutational diversity in many mtDNA regions. After adjustments of important sociodemographic and clinical variables, m.1005T>C, m.1824T>C, m.3394T>C, m.4491G>A, m.7828A>G, m.9814T>C, m.10586G>A, m.12338T>C, m.13708G>A, and m.14308T>C (at the Bonferroni-corrected significance) were negatively associated with short-term CD4+T-cell recovery whereas m.93A>G, m.15218A>G, and m.16399A>G were positively associated with short-term CD4+T-cell recovery.ConclusionOur baseline mtDNA characterization stresses the attention to East-Asian female PLWH at risk of CD4+T-cell loss-related aging and noncommunicable chronic diseases. Furthermore, mtDNA variants identified in regression analyses account for heterogeneity in short-term CD4+T-cell recovery of East-Asian PLWH. These results may help individualize the East-Asian immune recovery strategies under complicated HIV management caused by CD4+T-cell loss.

Immunological and virological response to initial antiretroviral therapy among older people living with HIV in the Canadian Observational Cohort (CANOC).

The aim of this study was to assess the adequacy of immunological recovery and virological suppression in response to antiretroviral therapy (ART) in the growing population of older people living with HIV (PLWH), as treatment regimens become more effective and tolerable. An interprovincial Canadian cohort of treatment-naïve PLWH who initiated ART after 1 January 2000 was used and age assessed in decades. Longitudinal absolute CD4 count response to treatment was modelled using generalized estimating equations. Cumulative incidence functions and proportional hazards models with a competing risk of death were used to estimate time to: (1) CD4≥200 cells/µL, (2) CD4≥500 cells/µL, (3) virological suppression (≤50 copies/mL), and (4) virological failure (>200 copies/mL). In all, 12489 individuals starting ART between 2000 and 2016 with one or more post-treatment CD4 count or viral load were included in the analysis. Age >60 years was associated with lower absolute CD4 recovery (adjusted β=-31 cells/µL) compared with age ≤30 years when pre-treatment CD4 count and other covariates were accounted for. Older age groups were less likely to achieve a CD4≥500 cells/µL, with the greatest effect in the >60 group [adjusted hazard ratio (aHR)=0.69, 95% confidence interval (CI): 0.57-0.84 vs. age ≤30). Older age groups were more likely to achieve viral suppression (age >60, aHR=1.20, 95% CI: 1.05-1.37) and less likely to have virological failure (age >60, aHR=0.46, 95% CI: 0.3-0.71) compared with those aged ≤30 years. Older adults have robust virological responses to ART; however, individuals over the age 60 are more likely to experience blunted CD4 recovery.

Differences in pyroptosis of recent thymic emigrants CD4+ T Lymphocytes in ART-treated HIV-positive patients are influenced by sex.

Pyroptosis cell death in recent thymus emigrants (RTE) CD4+ T lymphocytes plays an important role on HIV-1 infection as a cause of CD4+ T cell depletion, being influenced by several factors, among them, the sex. Thus, the aim of this study was evaluated pyroptosis levels in RTE CD4+ T lymphocytes of individuals under antiretroviral therapy (ART) stratified by sex. Thirty-seven ART-treated HIV-positive patients (22 females and 15 males) and 12 (seven females and five males) clinically health subjects were recruited. Analysis by flow-cytometry of RTE CD4+ cells (CD4+ CD31+ /fluorescent-labeled inhibitors of caspases-Caspase-1+) were performed. Clinical and sociodemographic aspects were also evaluated from medical records. We observed statistically higher levels of pyroptosis RTE CD4+ T cells in male individuals (69.3%) compared with female group (39.1%) (P=0.0356). Pre- and post-treatment CD4+ T cell counts were also higher in women than men (P=0.004 and P=0.012, respectively). Our data provides important evidence of the sex as a potential predictor of immunological reconstitution in ART-treated individuals.

Factors Impacting Differential Outcomes in the Definitive Radiation Treatment of Anal Cancer Between HIV-Positive and HIV-Negative Patients.

Anal squamous cell carcinoma (ASCC) is uncommon, yet seen more frequently in the setting of the human immunodeficiency virus (HIV). Chemoradiotherapy is the definitive modality of treatment for patients with ASCC; this study examines factors impacting clinical outcomes in a large cohort of HIV-positive and HIV-negative patients. A retrospective review was conducted of patients treated for nonmetastatic ASCC at a single institution between 2005 and 2018. Freedom from local recurrence (FFLR), freedom from distant metastasis, and overall survival (OS) were calculated using the Kaplan-Meier method, and univariate and multivariate analysis were performed using the Cox proportional hazards model. During the study period, 111 patients initiated definitive treatment for ASCC. Median age of the entire cohort was 56.7 years (interquartile range, 51.5-63.5), with 52 patients (46.8%) being HIV-positive. At median follow-up of 28.0 months, the 2- and 5-year FFLR were 78.2% (95% confidence interval [CI], 70.4-87.0) and 74.6% (95% CI, 65.8-84.5), respectively. Multivariate analysis revealed time from diagnosis to treatment initiation (median, 8 weeks; hazard ratio, 1.06; 95% CI, 1.03-1.10) to be significantly associated with worse FFLR and OS. HIV-positive patients had a trend toward worse FFLR (log-ranked p = .06). For HIV-positive patients with post-treatment CD4 less than 150 cells per mm3 , there was significantly worse OS (log-ranked p = .015). A trend toward worse FFLR was seen in HIV-positive patients, despite similar baseline disease characteristics as HIV-negative patients. Worse FFLR and OS was significantly associated with increased time from diagnosis to treatment initiation. Poorer OS was seen in HIV-positive patients with a post-treatment CD4 count less than 150 cells per mm3 . Human immunodeficiency virus (HIV)-positive patients with anal squamous cell carcinoma can represent a difficult clinical scenario. Definitive radiation with concurrent chemotherapy is highly effective but can result in significant toxicity and a decrease in CD4 count that could predispose to HIV-related complications. As HIV-positive patients have largely been excluded from prospective clinical trials, this study seeks to provide greater understanding of their outcomes with radiation therapy, potential predictors of worse local control and overall survival, and those most at risk after completion of treatment.

Immune Status and Associated Mortality After Cancer Treatment Among Individuals With HIV in the Antiretroviral Therapy Era

Immunologic decline associated with cancer treatment in people with HIV is not well characterized. Quantifying excess mortality associated with cancer treatment-related immunosuppression may help inform cancer treatment guidelines for persons with HIV. To estimate the association between cancer treatment and CD4 count and HIV RNA level in persons with HIV and between posttreatment CD4 count and HIV RNA trajectories and all-cause mortality. This observational cohort study included 196 adults with HIV who had an incident first cancer and available cancer treatment data while in the care of The Johns Hopkins HIV Clinic from January 1, 1997, through March 1, 2016. The study hypothesized that chemotherapy and/or radiotherapy in people with HIV would increase HIV RNA levels owing to treatment tolerability issues and would be associated with a larger initial decline in CD4 count and slower CD4 recovery compared with surgery or other treatment. An additional hypothesis was that these CD4 count declines would be associated with higher mortality independent of baseline CD4 count, antiretroviral therapy use, and risk due to the underlying cancer. Data were analyzed from December 1, 2017, through April 1, 2018. Initial cancer treatment category (chemotherapy and/or radiotherapy vs surgery or other treatment). Post-cancer treatment longitudinal CD4 count, longitudinal HIV RNA level, and all-cause mortality. Among the 196 participants (135 [68.9%] male; median age, 50 [interquartile range, 43-55] years), chemotherapy and/or radiotherapy decreased initial CD4 count by 203 cells/μL (95% CI, 92-306 cells/μL) among those with a baseline CD4 count of greater than 500 cells/μL. The decline for those with a baseline CD4 count of no greater than 350 cells/μL was 45 cells/μL (interaction estimate, 158 cells/μL; 95% CI, 31-276 cells/μL). Chemotherapy and/or radiotherapy had no detrimental association with HIV RNA levels. After initial cancer treatment, every 100 cells/μL decrease in CD4 count resulted in a 27% increase in mortality (hazard ratio, 1.27; 95% CI, 1.08-1.53), adjusting for HIV RNA level. No significant increase in mortality was associated with a unit increase in log10 HIV RNA after adjusting for CD4 count (hazard ratio, 1.24; 95% CI, 0.94-1.65). In this study, chemotherapy and/or radiotherapy was associated with significantly reduced initial CD4 count in adults with HIV compared with surgery or other treatment. Lower CD4 count after cancer treatment was associated with an increased hazard of mortality. Further research is necessary on the immunosuppressive effects of cancer treatment in adults with HIV and whether health care professionals must consider the balance of cancer treatment efficacy against the potential cost of further immunosuppression. Monitoring of immune status may also be helpful given the decrease in CD4 count after treatment and the already immunocompromised state of patients with HIV.

Abstract LB-223: ATLASTMreveals a dominant inhibitory antigen in melanoma patients, and a reduced breadth of tumor-associated antigen-specific T cells in non-responders to checkpoint blockade

Abstract Broadly reinvigorating the T cell response by blocking immune-inhibitory receptors can produce durable clinical benefit in 20-40% of melanoma patients. Due to low response rates and potential toxicity of checkpoint therapy, identification of biomarkers to predict response to immune checkpoint inhibitors (ICIs) has the potential to stratify patients and increase the efficacy of ICI therapy in treated patients. However, to date, the significant effort to identify biomarkers has yielded unreliable predictors. To better understand ICI response and resistance, we profiled T cell responses to a panel of known tumor-associated antigens (TAAs). The ex vivo ATLAS™ technology enables comprehensive profiling of a patient’s antigen-specific T cells, with putative antigens expressed as individual clones, processed by the subject’s own antigen presenting cells and presented to autologous CD4+ or CD8+ T cells for measurement of recall responses. A unique feature of ATLAS is the ability to identify stimulatory and inhibitory T cell responses based on cytokine secretion that statistically exceeds or is reduced, respectively, relative to baseline controls. Pre- and post-treatment CD4+ and CD8+ T cell responses to a library of 56 known TAAs were profiled from 16 patients with metastatic melanoma undergoing ICI therapy. Consistent with previous studies, a low frequency of responses was observed from both T cell subsets. Overall, there were fewer responses detected in non-responders than in responders both pre- and post-treatment; in fact, no stimulatory CD8+ T cell responses were identified in non-responders prior to ICI therapy. Surprisingly, one TAA elicited only inhibitory responses across most subjects, independently of clinical outcome and predominantly from the CD8+ T cell subset. Hierarchical clustering showed that patient responses are similar to themselves across visits, but the antigen profiles across patients did not cluster by visit or treatment response. ATLAS provides a powerful, non-invasive, blood-based platform for the interrogation of peripheral T cell responses. Our data show that melanoma patients who are non-responsive to ICI therapy have a limited T cell response to TAAs. In addition, one TAA elicited a dominant inhibitory response across patients. Upon further refinement, patient-specific responses may allow for prediction of advanced melanoma patients for whom checkpoint therapy would prove successful. Citation Format: Louisa Dowal, Anna Lyubetskaya, Crystal Cabral, Mariya Croll, Christopher Warren, James Perry, Melissa Hayes, James Loizeaux, James Foti, Jessica B. Flechtner, Jason R. Dobson, F. Stephen Hodi, Wendy Broom. ATLASTMreveals a dominant inhibitory antigen in melanoma patients, and a reduced breadth of tumor-associated antigen-specific T cells in non-responders to checkpoint blockade [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr LB-223.

Factors affecting differential outcomes in the definitive treatment of anal cancer between HIV+ and HIV- patients.

3572 Background: Anal cancer is an uncommon malignancy with numerous factors that influence treatment outcomes. Historically, HIV+ patients were restricted from entering clinical trials, limiting data on their outcomes to small retrospective reports. This study seeks to understand the factors related to anal cancer outcomes, specifically the differences between HIV+ and HIV- patients. Methods: Inclusion criteria was non-metastatic anal squamous cell carcinoma treated with a definitive course of chemotherapy and radiation between 2005 and 2018 at a single institution. Clinical data related to baseline characteristics, treatment parameters, and post-treatment follow-up were extracted for calculation of freedom from local recurrence (FFLR) and overall survival (OS). Univariate analysis (UVA) and multivariate analysis (MVA) were done using cox proportional hazard model, and FFLR and OS were calculated using the Kaplan-Meier method. Results: During the study period, 111 patient initiated definitive treatment for anal cancer. Median age was 56.7 years (IQR: 51.4-63.5), and 47% (N = 52) were HIV+. At median follow-up of 28 months, 12 and 24-month FFLR was 84.1% and 78.2% respectively, with 24-month OS of 87.3%. MVA demonstrated significant association between FFLR and T-stage HR 4.02 (95% CI: 2.14-7.55) p &lt; 0.001, elapsed treatment time (median of 50 days) 1.08 (95% CI: 1.04-1.12) p &lt; 0.001, and diagnosis to treatment start (median time of 15 weeks) 1.05 (95% CI: 1.01-1.08) p = 0.005. Additional analysis with log-rank test for FFLR demonstrated significant difference between patients taking &lt; 50 days to complete treatment (p = 0.03), and &lt; 15 weeks from diagnosis to treatment completion (p = 0.006). In HIV+ patients, post-treatment CD4 &lt; 150 was significantly associated with worse OS on log-rank test (p = 0.016), with pretreatment CD4 values being non-significant. Conclusions: This study represents the largest single institution report of HIV positive patients treated for anal cancer. No difference in local recurrence or overall survival between HIV+ and HIV- patients was elucidated; however, HIV+ patients with lower post-treatment CD4 counts had worse OS. The most significant predictors of local recurrence were advanced T-stage, increased time from diagnosis to treatment initiation, and prolonged treatment time.

Nano-Pulse Stimulation Ablates Orthotopic Rat Hepatocellular Carcinoma and Induces Innate and Adaptive Memory Immune Mechanisms that Prevent Recurrence.

Nano-pulse stimulation (NPS), previously called nsPEFs, induced a vaccine-like effect after ablation of orthotopic N1-S1 hepatocellular carcinoma (HCC), protecting rats from subsequent challenges with N1-S1 cells. To determine immunity, immune cell phenotypes were analyzed in naïve, treated and protected rats. NPS provides a positive, post-ablation immuno-therapeutic outcome by alleviating immunosuppressive T regulatory cells (Treg) in the tumor microenvironment (TME), allowing dendritic cell influx and inducing dynamic changes in natural killer cells (NKs), NKT-cells and T-lymphocytes in blood, spleen and liver. NPS induced specific increases in NKs and NKT-cells expressing CD8 and activation receptors CD314-NKG2D and CD161 (NK1.1) in the TME after treatment, as well as some variable changes in CD4+ and CD8+ effector (Tem) and central memory (Tem) lymphocytes in blood and spleen. After orthotopic challenge, CD8+ T-cells were cytotoxic, inducing apoptosis in N1-S1 cells; additionally, in contrast to post-treatment immune responses, CD4+ and CD8+ memory precursor effector cells (MPECs) and short-lived effector cells (SLECs) were present, while still including CD8+ CD161 NK cells, but not involving CD8+ CD314-NKG2D+ NKs. This immunity was N1-S1-specific and was sustained for at least 8 months. NPS vaccinates rats in vivo against HCC by activating innate and adaptive immune memory mechanisms that prevent HCC recurrence.

Early Stage HER2-Positive Breast Cancers Not Achieving a pCR From Neoadjuvant Trastuzumab- or Pertuzumab-Based Regimens Have an Immunosuppressive Phenotype

BackgroundStromal tumor-infiltrating lymphocytes (TILs) might predict pathologic complete response (pCR) in patients with HER2-positive (HER2+) breast cancer treated with trastuzumab (H). Docetaxel (T), carboplatin (C), H, and pertuzumab (P) have immune-modulating effects. Pre- and post-treatment immune biomarkers in cancers treated with neoadjuvant TCH with or without P are lacking. In this study we quantified baseline and changes in TILs, cluster of differentiation (CD) 4+, CD8+, FoxP3+, and PD-L1+ cells using immunohistochemistry (IHC) and quantified productive T-cell receptor β (TCRβ) rearrangements and TCRβ clonality using next-generation sequencing (NGS) in 30 HER2+ breast cancer tissues treated with neoadjuvant H with or without P regimens. Materials and MethodsThirty pre- and post-neoadjuvant TCH (n = 4) or TCHP (n = 26) breast cancer tissues were identified. TILs were quantified manually using hematoxylin and eosin. CD4, CD8, FoxP3, and PD-L1 were stained using IHC. TCRβ was evaluated using NGS. Immune infiltrates were compared between pCR and non-pCR groups using the Wilcoxon rank sum test. ResultsA pCR occurred in 15 (n = 15; 50%) cancers (TCH n = 2; TCHP, n = 13). Pretreatment TILs, CD4+, CD8+, FoxP3+, and PD-L1+ cells were not associated with response (P = .42, P = .55, P = .19, P = .66, P = .87, respectively. Pretreatment productive TCRβ and TCRβ clonality did not predict response, P = .84 and P = .40, respectively). However, post-treatment CD4+ and FoxP3+ cells (T-regulatory cells) were elevated in the non-pCR cohort (P = .042 and P = .082, respectively). ConclusionAn increase in regulatory T cells in non-pCR tissues suggests the development of an immunosuppressive phenotype. Further investigation in a larger cohort of samples is warranted to validate these findings.

Impact of Posttreatment CD4 Count on Tumor Recurrence Among HIV Positive Anal Cancer Patients

Impact of Posttreatment CD4 Count on Tumor Recurrence Among HIV Positive Anal Cancer Patients

Effect of CD4 Count on Treatment Toxicity and Tumor Recurrence in Human Immunodeficiency Virus–Positive Patients With Anal Cancer

Tostudy the effects of immunosuppression on treatment toxicity, long-term cancer recurrence risk, and survival among human immunodeficiency virus (HIV)-positive anal cancer patients. From a nationwide retrospective cohort of veterans with anal cancer we identified 142 HIV-positive patients with stage I-III disease, diagnosed between 2000 and 2015 and treated with definitive-intent chemotherapy and radiation. We used regression models to study the impact of pretreatment CD4 counts and longitudinal posttreatment CD4 counts on outcomes including acute toxicity, long-term ostomy rates, cancer recurrence, cancer-specific survival, and overall survival. All models were adjusted for potential confounders. The median pretreatment CD4 count was 375cells/mm3, which dropped to 157cells/mm3 after treatment. Each 100-cell/mm3 decrease in pretreatment CD4 count was associated with an increased risk of acute hematologic toxicity (odds ratio 1.19, 95% confidence interval [CI] 1.01-1.42, P=.04) and hospitalization for hematologic toxicity (odds ratio 1.24, 95% CI 1.00-1.54, P=.049) but not gastrointestinal toxicity, tumor recurrence, or cancer-specific mortality (P>.05). Each 100-cells/mm3 decrease in posttreatment CD4 count increased the risk of recurrence by 54% (hazard ratio 1.54, 95% CI 1.09-2.17, P=.01) and cancer mortality by 46% at a trend level (hazard ratio 1.46, 95% CI 0.99-2.14, P=.06). Neither pre- nor posttreatment CD4 count influenced long-term ostomy rates or overall survival (all P>.05). Lower pretreatment CD4 counts were associated with acute hematologic toxicity, and lower posttreatment CD4 count levels were associated with an increased risk of tumor recurrence. These results suggest that immune surveillance may play an important role in long-term disease control in anal cancer.

Abstract 1611: Intravesical BCG induces CD4 T Cell expansion in an immune competent model of bladder cancer

Abstract Intravesical BCG Immunotherapy is the standard of care in treating non-muscle invasive bladder cancer (NMIBC), yet its mechanism of action remains elusive. Both innate and adaptive immune responses have been implicated in BCG activity. While prior research has indirectly demonstrated the importance of T cells and shown a rise in CD4+ T cells in bladder tissue after BCG, T cell subpopulations have not been fully characterized. We investigated the relationship between effector and regulatory T cells in an immune competent, clinically relevant rodent model of bladder cancer. Fischer 344 rats aged 7 weeks received 1.5mg/kg N-Nitroso-N-methylurea (MNU) every other week for 6 weeks (4 doses). Bladder dysplasia began by week 8 and by week 16 the majority of rats had a NMIBC phenotype. Beginning week 8 following the first MNU dose, rats were intravesically administered 0.3ml of BCG (Tice), cisplatin (1mg/ml), Mitomycin C (2mg/ml), MMC+ BCG, or saline (n=10 for all groups) weekly for 6 total doses. Animals were sacrificed at week 16, and bladders were processed for histopathology and digested into single cell suspensions for flow cytometry. Whole transcriptome expression profiling was then performed on sorted CD4 and CD8 cells of post-BCG tumors vs untreated tumors to assess T cell differentiation after BCG. Our data demonstrate that cancer progression in the MNU rat model of bladder cancer is characterized by a decline in the CD8/FoxP3 ratio, consistent with decreased adaptive immunity. By contrast, treatment with intravesical BCG leads to a large, transient rise in the CD4+ T cell population in the urothelium, and is both more effective and immunogenic compared to intravesical chemotherapy. Interestingly, whole transcriptome expression profiling of post-treatment intravesical CD4+ and CD8+ T cells revealed minimal differences in gene expression after BCG treatment. Together, our results suggest that while BCG induces T cell recruitment to the bladder, the T cell phenotype does not markedly change, implying that combining T cell activating agents with BCG might improve clinical activity. Citation Format: Max Kates, Thomas Nirschl, Nikolai Sopko, Noah Hahn, David McConkey, Alex Baras, Charles Drake, Trintiy Bivalacqua. Intravesical BCG induces CD4 T Cell expansion in an immune competent model of bladder cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 1611. doi:10.1158/1538-7445.AM2017-1611