The HIV program in Newfoundland and Labrador (NL) provides care for all persons living with HIV (PLWH) in NL, yet progress toward UNAIDS 95-95-95 goals for diagnosis, linkage to care and viral suppression has not previously been documented. This analysis describes engagement in HIV care and virologic outcomes for the NL cohort in 2016 and 2019 and compares this data to the Canadian HIV Observational Cohort (CANOC). A retrospective review of the NL clinic included adults aged >18 years and descriptive statistics for demographics, risk factors, and clinical variables were assessed and compared using χ2 test or Fisher's Exact test (categorical) or Wilcoxon Sum Rank test (continuous). Engagement in care and virologic outcomes for the NL cohort were consistently high over the 2016 to 2019 period with > 98% on antiretroviral therapy (ART), and > 96% having a suppressed virus load. Engagement in care and virologic outcomes among PLWH in NL is high and compares favorably to a national cohort.

Natural killer (NK) and CD8+ T cell function is compromised in human immunodeficiency virus type 1 (HIV-1) infection by increased expression of inhibitory receptors such as TIGIT (T cell immunoreceptor with Ig and ITIM domains). Blocking inhibitory receptors or their ligands with monoclonal antibodies (mAb) has potential to improve antiviral immunity in general and facilitate HIV eradication strategies. We assessed the impact of TIGIT engagement and blockade on cytotoxicity, degranulation, and interferon-gamma (IFN-γ) production by CD8+ T cells from persons living with HIV (PLWH). The effect of TIGIT engagement on non-specific anti-CD3-redirected cytotoxicity was assessed in redirected cytotoxicity assays, and the effect of TIGIT blockade on HIV-specific CD8+ T cell responses was assessed by flow cytometry. In 14/19 cases where peripheral blood mononuclear cells (PBMC) mediated >10% redirected cytotoxicity, TIGIT engagement reduced the level of cytotoxicity to <90% of control values. We selected PLWH with >1000 HIV Gag or Nef-specific IFN-γ spot forming cells per million PBMC to quantify the effects of TIGIT blockade on HIV-specific CD8+ T cell responses by flow cytometry. Cell surface TIGIT expression decreased on CD8+ T cells from 23/40 PLWH following TIGIT blockade and this loss was associated with increased anti-TIGIT mAb fluorescence on monocytes. In total, 6 of these 23 PLWH had enhanced HIV-specific CD8+ T cell degranulation and IFN-γ production with TIGIT blockade, compared to 0/17 with no decrease in cell surface TIGIT expression. Reduced CD8+ T cell TIGIT expression with TIGIT blockade involved trogocytosis by circulating monocytes, suggesting that an effector monocyte population and intact fragment crystallizable (Fc) functions are required for mAb-based TIGIT blockade to effectively enhance HIV-specific CD8+ T cell responses.

Infection before primary vaccination (herein termed “hybrid immunity”) engenders robust humoral immunity and broad Ab-dependent cell-mediated cytotoxicity (ADCC) across SARS-CoV-2 variants. We measured and compared plasma IgG and IgA against Wuhan-Hu-1 and Omicron (B.1.1.529) full-length spike (FLS) and receptor binding domain after three mRNA vaccines encoding Wuhan-Hu-1 spike (S) and after Omicron breakthrough infection. We also measured IgG binding to Wuhan-Hu-1 and Omicron S1, Wuhan-Hu-1 S2 and Wuhan-Hu-1 and Omicron cell-based S. We compared ADCC using human embryonic lung fibroblast (MRC-5) cells expressing Wuhan-Hu-1 or Omicron S. The effect of Omicron breakthrough infection on IgG anti-Wuhan-Hu-1 and Omicron FLS avidity was also considered. Despite Omicron breakthrough infection increasing IgG and IgA against FLS and receptor binding domain to levels similar to those seen with hybrid immunity, there was no boost to ADCC. Preferential recognition of Wuhan-Hu-1 persisted following Omicron breakthrough infection, which increased IgG avidity against Wuhan-Hu-1 FLS. Despite similar total anti-FLS IgG levels following breakthrough infection, 4-fold higher plasma concentrations were required to elicit ADCC comparable to that elicited by hybrid immunity. The greater capacity for hybrid immunity to elicit ADCC was associated with a differential IgG reactivity pattern against S1, S2, and linear determinants throughout FLS. Immunity against SARS-CoV-2 following Omicron breakthrough infection manifests significantly less ADCC capacity than hybrid immunity. Thus, the sequence of antigenic exposure by infection versus vaccination and other factors such as severity of infection affect antiviral functions of humoral immunity in the absence of overt quantitative differences in the humoral response.

Some SARS-CoV-2-exposed individuals develop immunity without overt infection. We identified 11 individuals who were negative by nucleic acid testing during prolonged close contact and with no serological diagnosis of infection. As this could reflect natural immunity, cross-reactive immunity from previous coronavirus exposure, abortive infection due to de novo immune responses, or other factors, our objective was to characterize immunity against SARS-CoV-2 in these individuals. Blood was processed into plasma and peripheral blood mononuclear cells (PBMC) and screened for IgG, IgA, and IgM antibodies (Ab) against SARS-CoV-2 and common β-coronaviruses OC43 and HKU1. Receptor blocking activity and interferon-alpha (IFN-α) in plasma were also measured. Circulating T cells against SARS-CoV-2 were enumerated and CD4+ and CD8+ T cell responses discriminated after in vitro stimulation. Exposed uninfected individuals were seronegative against SARS-CoV-2 spike (S) and selectively reactive against OC43 nucleocapsid protein (N), suggesting common β-coronavirus exposure induced Ab cross-reactive against SARS-CoV-2 N. There was no evidence of protection from circulating angiotensin-converting enzyme (ACE2) or IFN-α. Six individuals had T cell responses against SARS-CoV-2, with four involving CD4+ and CD8+ T cells. We found no evidence of protection from SARS-CoV-2 through innate immunity or immunity induced by common β-coronaviruses. Cellular immune responses against SARS-CoV-2 were associated with time since exposure, suggesting that rapid cellular responses may contain SARS-CoV-2 infection below the thresholds required for a humoral response.

Mucosal IgA is widely accepted as providing protection against respiratory infections, but stimulation of mucosal immunity, collection of mucosal samples and measurement of mucosal IgA can be problematic. The relationship between mucosal and circulating IgA responses is unclear, however, whole blood is readily collected and circulating antigen-specific IgA easily measured. We measured circulating IgA against SARS-CoV-2 spike (S) to investigate vaccine- and infection-induced production and correlation with protection. Circulating IgA against ancestral (Wuhan-Hu-1) and Omicron (BA.1) S proteins was measured at different time points in a total of 143 subjects with varied backgrounds of vaccination and infection. Intramuscular vaccination induced circulating anti-SARS-CoV-2 S IgA. Subjects with higher levels of vaccine-induced IgA against SARS-CoV-2 S (p = 0.0333) or receptor binding domain (RBD) (p = 0.0266) were less likely to experience an Omicron breakthrough infection. The same associations did not hold for circulating IgG anti-SARS-CoV-2 S levels. Breakthrough infection following two vaccinations generated stronger IgA anti-SARS-CoV-2 S responses (p = 0.0002) than third vaccinations but did not selectively increase circulating IgA against Omicron over ancestral S, indicating immune imprinting of circulating IgA responses. Circulating IgA against SARS-CoV-2 S following breakthrough infection remained higher than vaccine-induced levels for over 150 days. In conclusion, intramuscular mRNA vaccination induces circulating IgA against SARS-CoV-2 S, and higher levels are associated with protection from breakthrough infection. Vaccination with ancestral S enacts imprinting within circulating IgA responses that become apparent after breakthrough infection with Omicron. Breakthrough infection generates stronger and more durable circulating IgA responses against SARS-CoV-2 S than vaccination alone.

Tungsten is widely used in medical, industrial, and military applications. The environmental exposure to tungsten has increased over the past several years, and few studies have addressed its potential toxicity. In this study, we evaluated the effects of chronic oral tungsten exposure (100 ppm) on renal inflammation in male mice. We found that 30- or 90-day tungsten exposure led to the accumulation of LAMP1-positive lysosomes in renal tubular epithelial cells. In addition, the kidneys of mice exposed to tungsten showed interstitial infiltration of leukocytes, myeloid cells, and macrophages together with increased levels of proinflammatory cytokines and p50/p65-NFkB subunits. In proximal tubule epithelial cells (HK-2) in vitro, tungsten induced a similar inflammatory status characterized by increased mRNA levels of CSF1, IL34, CXCL2, and CXCL10 and NFkB activation. Moreover, tungsten exposure reduced HK-2 cell viability and enhanced reactive oxygen species generation. Conditioned media from HK-2 cells treated with tungsten induced an M1-proinflammatory polarization of RAW macrophages as evidenced by increased levels of iNOS and interleukin-6 and decreased levels of the M2-antiinflammatory marker CD206. These effects were not observed when RAW cells were exposed to conditioned media from HK-2 cells treated with tungsten and supplemented with the antioxidant N-acetylcysteine (NAC). Similarly, direct tungsten exposure induced M1-proinflammatory polarization of RAW cells that was prevented by NAC co-treatment. Altogether, our data suggest that prolonged tungsten exposure leads to oxidative injury in the kidney ultimately leading to chronic renal inflammation characterized by a proinflammatory status in kidney tubular epithelial cells and immune cell infiltration.

Antibodies capable of neutralizing SARS-CoV-2 are well studied, but Fc receptor–dependent antibody activities that can also significantly impact the course of infection have not been studied in such depth. Since most SARS-CoV-2 vaccines induce only anti-spike antibodies, here we investigated spike-specific antibody-dependent cellular cytotoxicity (ADCC). Vaccination produced antibodies that weakly induced ADCC; however, antibodies from individuals who were infected prior to vaccination (hybrid immunity) elicited strong anti-spike ADCC. Quantitative and qualitative aspects of humoral immunity contributed to this capability, with infection skewing IgG antibody production toward S2, vaccination skewing toward S1, and hybrid immunity evoking strong responses against both domains. A combination of antibodies targeting both spike domains support strong antibody-dependent NK cell activation, with 3 regions of antibody reactivity outside the receptor-binding domain (RBD) corresponding with potent anti-spike ADCC. Consequently, ADCC induced by hybrid immunity with ancestral antigen was conserved against variants containing neutralization escape mutations in the RBD. Induction of antibodies recognizing a broad range of spike epitopes and eliciting strong and durable ADCC may partially explain why hybrid immunity provides superior protection against infection and disease compared with vaccination alone, and it demonstrates that spike-only subunit vaccines would benefit from strategies that induce combined anti-S1 and anti-S2 antibody responses.

Objective To assess the incidence of COVID-19 infection in the absence of a confirmatory test in persons suspecting they contracted COVID-19 and elucidate reasons for their belief. Methods We recruited persons with a confirmed COVID-19 diagnosis and persons who believed they may have contracted COVID-19 between December, 2019 and April, 2021 into a study of immunity against SARS-CoV-2. An intake questionnaire captured their perceived risk factors for exposure and symptoms experienced, including symptom duration and severity. ELISA testing against multiple SARS-CoV-2 antigens was done to detect antibodies against SARS-CoV-2. No participant had received COVID-19 vaccination prior to the time of testing. Results The vast majority of study subjects without Public Health confirmation of infection had no detectable antibodies against SARS-CoV-2. Suspected infection with SARS-CoV-2 generally involved experiencing symptoms common to many other respiratory infections. Unusually severe or persistent symptoms often supported suspicion of infection with SARS-CoV-2 as did travel or contact with travelers from outside Newfoundland and Labrador. Rare cases in which antibodies against SARS-CoV-2 were detected despite negative results of Public Health testing for SARS-CoV-2 RNA involved persons in close contact with confirmed cases. Conclusions Broad public awareness and declaration of pandemic status in March, 2020 contributed to the perceived risk of contracting COVID-19 in Newfoundland and Labrador from late 2019 to April 2021 and raised expectation of its severity. Serological testing is useful to diagnose past infection with SARS-CoV-2 to accurately estimate population exposure rates.

Extensive inflammation in the liver is known to contribute to the pathogenesis of hepatitis C virus (HCV) infection. Apoptosis has, for a long time, been known to act as a mechanism of hepatocyte death, but our previous research also identified inflammasome-mediated pyroptosis in infected and uninfected bystander cells as an additional mechanism of HCV-induced cytopathicity. The purpose of this study was to investigate the mechanism of HCV-induced cell death and to determine the timing and relative contributions of apoptosis and pyroptosis during HCV infection. In a model employing a cell culture-adapted strain of JFH-1 HCV and Huh-7.5 hepatocyte-like cells, we found that pyroptosis occurred earlier than did apoptosis during infection. CRISPR knockout of NLRP3 resulted in decreased caspase-1 activation, but not complete elimination, indicating multiple sensors are likely involved in HCV-induced pyroptosis. Knockout of gasdermin-D resulted in increased activation of apoptosis-related caspase-3, suggesting potential crosstalk between the two cell death pathways. An unexpected decrease in activated caspase-1 levels was observed when caspase-3 was knocked out, implying that caspase-3 may have a role in the initiation of pyroptosis, at least in the context of HCV infection. Lower viral titres in culture fluids and increased ratios of intracellular to extracellular levels of infectious virus were observed in knockout versus wild-type Huh-7.5 cells, suggesting that HCV may induce programmed cell death in order to enhance virus release from infected cells. These results contribute to the understanding of HCV pathogenesis and add to the increasing volume of literature suggesting various programmed cell death pathways are not mutually exclusive.

Despite improvements in outcomes for patients with EGFR-mutant NSCLC, tumors inevitably relapse during tyrosine kinase inhibitor (TKI) treatment. Alterations in tumor suppressor genes (TSGs) have been described as determinants of molecular and clinical heterogeneity in EGFR-mutant NSCLC, however, detailed outcome analyses based on specific TSGs are limited.

Background Immune checkpoint inhibitors (ICI) improve overall survival (OS) in patients with locally advanced, unresectable, or metastatic urothelial carcinoma (aUC), but response rates can be modest. We compared outcomes between patients with and without prior intravesical Bacillus Calmette-Guerin (BCG), who received ICI for aUC, hypothesizing that prior intravesical BCG would be associated with worse outcomes. Patients and Methods We performed a retrospective cohort study across 25 institutions in US and Europe. We compared observed response rate (ORR) using logistic regression; progression-free survival (PFS) and OS using Kaplan-Meier and Cox proportional hazards. Analyses were stratified by treatment line (first line/salvage) and included multivariable models adjusting for known prognostic factors. Results A total of 1026 patients with aUC were identified; 614, 617, and 638 were included in ORR, OS, PFS analyses, respectively. Overall, 150 pts had history of prior intravesical BCG treatment. ORR to ICI was similar between those with and without prior intravesical BCG exposure in both first line and salvage settings (adjusted odds radios 0.55 [P= .08] and 1.65 [P= .12]). OS (adjusted hazard ratios 1.05 [P= .79] and 1.13 [P= .49]) and PFS (adjusted hazard ratios 1.12 [P= .55] and 0.87 [P= .39]) were similar between those with and without intravesical BCG exposure in first line and salvage settings. Conclusion Prior intravesical BCG was not associated with differences in response and survival in patients with aUC treated with ICI. Limitations include retrospective nature, lack of randomization, presence of selection and confounding biases. This study provides important preliminary data that prior intravesical BCG exposure may not impact ICI efficacy in aUC.

Viral ImmunologyVol. 35, No. 5 EditorialFree AccessThe Value of “00” OneMichael D. Grant and Rodney S. RussellMichael D. GrantAddress correspondence to: Dr. Michael D. Grant, BioMedical Sciences, Memorial University of Newfoundland, 300 Prince Philip Dr., St. John's, Newfoundland and Labrador, A1B 3V6, Canada E-mail Address: mgrant@mun.caImmunology and Infectious Diseases Program, Division of BioMedical Sciences, Faculty of Medicine, Memorial University of Newfoundland and Labrador, St. John's, NL, Canada.Search for more papers by this author and Rodney S. RussellImmunology and Infectious Diseases Program, Division of BioMedical Sciences, Faculty of Medicine, Memorial University of Newfoundland and Labrador, St. John's, NL, Canada.Search for more papers by this authorPublished Online:14 Jun 2022https://doi.org/10.1089/vim.2022.0087AboutSectionsPDF/EPUB Permissions & CitationsPermissionsDownload CitationsTrack CitationsAdd to favorites Back To Publication ShareShare onFacebookTwitterLinked InRedditEmail For any study involving human subjects, we acknowledge the study participants for their willingness to provide specimens and sometimes very personal medical details that allow us to carry out the research we perform. We refer to these individuals as study subjects, participants, patients, cohorts, for example, but these are people. In an effort to de-identify them, perhaps we somewhat de-humanize them when we tell their stories. These individuals are people, people we may have met, people who are often sick, often vulnerable. These people have entered an agreement of trust with us, often for very little or no personal gain, that directly enables us to do the research we want to do.In recent years, there has been a shift toward involving the study participants more and more in the research. These days, study participants even can be involved in the planning of the research from the start. This is particularly important when study populations involve vulnerable groups. We try to do research “with people” instead of “on them.” In many cases, principal investigators may never meet the people who provide specimens or scientific data, but this is not always the case. In other studies, enrollment into a study has translated to decades of collaboration and personal interaction where the people in the study have gotten to know the researchers very well. The loss then that can be felt when a study subject passes can be as hard as losing a friend.Below is a tribute, written by my mentor, friend, and colleague, Dr. Michael Grant, about the first person he enrolled in his first research study almost 30 years ago.In July 1995, I started my independent research career in a new location with new expectations among new colleagues and staff. In November that year, we enrolled the first volunteer (subject 001) into a study on the immunology of human immunodeficiency virus (HIV) infection. Sadly, this May, the person I first knew as subject 001 passed away, prompting thoughtful reflection on the nature of our longstanding relationship, a relationship founded on their desire to help generate new knowledge for the greater good, while receiving nothing tangible in exchange. Outside of longitudinal aging studies, providing informed consent does not normally imply study participation lasting through three decades, but investigators such as myself can benefit tremendously from consistent long-term access to sequential samples from the same individuals.Over a period of 27 years, 001's plasma was pipetted into thousands of ELISA plates to release information on their evolving antibody repertoire, circulating cytokine milieu and immune activation status. Their plasma recorded a diverse and challenging history of antiretroviral drug treatment and was sourced for early drug resistance genotyping. It marked infected cells for targeting by natural killer (NK) cells and yielded viruses for sequence analysis. Their lymphocytes were equally valuable and informative. B lymphocytes were transformed into a permanent cell line for autologous targets of cytotoxic T cells (CTL) and T lymphocytes were screened for reactivity against recombinant viruses, viral peptides, and unknown antigens on a variety of targets. The adaptive phenotypic and functional properties of their T lymphocytes, dictated by aging with HIV infection, were captured in ELISPOT, cytotoxicity, and flow cytometry assays spanning 27 years.Trainees, staff, and colleagues came and went, while 001's samples flowed regularly to my lab, at widening intervals as antiretroviral therapy and HIV clinical care matured towards the standard enjoyed by people living with HIV (PLWH) today. The early days of treatment with the “D” drugs with their mitochondrial toxicity, followed by the lipodystrophy and lipoatrophy emerging during the time of protease inhibitors, took a toll on many PLWH. Increased incidence of type II diabetes, renal disease and accelerated aging emerged as new issues, but 001's resilience over that period symbolized that of their community. The transformation brought about by effective combination antiretroviral therapy is faithfully reflected in 001's laboratory records over their 99 visits. Their CD4+ T cell counts rebounded from 51/μL to 840/μL, plasma virus load fell from >500,000 to <30 copies/mL and plasma β2 microglobulin dropped from 11.3 μg/mL to 2.4 μg/mL. Through the many experiments enabled by sequential blood donations, 001 was a constant contributor to research publications and a part of almost all my grant proposals and presentations.My first graduate student used 001's CD8+ T cells to characterize autoreactivity against uninfected CD4+ T cells and to describe a novel form of activation-induced cell death (Kottilil et al., 2001; Kottilil et al., 2000). One of the first studies to link HIV reservoir size with immunological and clinical outcomes used 001's CD4+ T cells (Russell et al., 2001). Their CD8+ T cell's genomes helped to demonstrate T cell repertoire preservation despite falling numbers and the functional and phenotypic attributes of their CD8+ T cells outlined relationships between HIV disease progression, immune memory, immune reconstitution, antiretroviral treatment, and viral suppression (Gamberg et al., 2004a; Gamberg et al., 2004b; Gamberg et al., 1999; Mason et al., 2004).During the H1N1 influenza pandemic, their response to vaccination formed part of a data set illustrating issues relevant to PLWH vaccination (Kelly et al., 2012). Their HIV-specific CTL responded favorably to checkpoint inhibitors, helping to usher in a new era of ideas for immunotherapy and their NK cells were positioned by case history within a rare subset critical for addressing the role of NK cell adaptation in HIV infection (Adegoke et al., 2015; Holder et al., 2021).I have no doubt we all appreciate the efforts made by volunteer study subjects to contribute to our research, as without them, progress is literally impossible. However, while reflecting on the contributions 001 made over the past 27 years, I realized how much more useful, meaningful, and abundant those contributions were than I was generally aware. The gift that research study subjects give is seldom directly returned and they often remained silent partners supporting my progress and that of a growing series of trainees. I owe a debt of gratitude to all the volunteers who participated in our research studies as PLWH have always been enthusiastic participants, but the breadth of impact that 001 has had strikes home most deeply.I had the good fortune to meet subject 001 several times during my career and was struck by their broad compassion and gentle giving nature. The individual was an advocate for PLWH, an animal lover, a politician, and a craftsperson, and lived through the transformative era in HIV care, while willingly contributing more than was asked, touching myself, my colleagues, and many lab members in the process. Far exceeding the bounty of experimental data, I am left the lasting gift of the memory of a person who gave of themselves readily, extensively, and selflessly, and I learned more than I dared hope in ways I never expected. I take this opportunity to thank you and all the other volunteers contributing samples for scientific research. Your legacy is progress.ReferencesAdegoke A, Gladney K, Gallant M, et al. Heteroclitic Peptides Increase Proliferation and Reduce Evidence of Human Immunodeficiency Virus-Specific CD8(+) T Cell Dysfunction. Viral Immunol 2015;28:455–463. doi: 10.1089/vim.2015.0036. Link, Google ScholarGamberg J, Barrett L, Bowmer MI, et al. Factors related to loss of HIV-specific cytotoxic T lymphocyte activity. AIDS 2004a;18:597–604. doi: 10.1097/00002030-200403050-00003. Crossref, Medline, Google ScholarGamberg J, Pardoe I, Bowmer MI, et al. Lack of CD28 expression on HIV-specific cytotoxic T lymphocytes is associated with disease progression. Immunol Cell Biol 2004b;82:38–46. doi: 10.1111/j.1440-1711.2004.01204.x. Crossref, Medline, Google ScholarGamberg JC, Bowmer MI, Trahey JC, et al. Functional and genetic integrity of the CD8 T-cell repertoire in advanced HIV infection. AIDS 1999;13:2043–2053. doi: 10.1097/00002030-199910220-00006. Crossref, Medline, Google ScholarHolder KA, Burt K, Grant MD. TIGIT blockade enhances NK cell activity against autologous HIV-1-infected CD4(+) T cells. Clin Transl Immunol 2021;10:e1348. doi: 10.1002/cti2.1348. eCollection 2021. Crossref, Medline, Google ScholarKelly D, Burt K, Missaghi B, et al. Responses to pandemic ASO3-adjuvanted A/California/07/09 H1N1 influenza vaccine in human immunodeficiency virus-infected individuals. BMC Immunol 2012;13:49. doi: 10.1186/1471-2172-13-49. Crossref, Medline, Google ScholarKottilil S, Bowmer MI, Trahey J, et al. Fas/FasL-independent activation-induced cell death of T lymphocytes from HIV-infected individuals occurs without DNA fragmentation. Cell Immunol 2001;214:1–11. doi: 10.1006/cimm.2001.1876. Crossref, Medline, Google ScholarKottilil S, Gamberg J, Bowmer I, et al. Human immunodeficiency virus type 1 replication, immune activation, and circulating cytotoxic T cells against uninfected CD4+ T cells. J Clin Immunol 2000;20:175–186. doi: 10.1023/a:1006633429087. Crossref, Medline, Google ScholarMason RD, Bowmer MI, Howley CM, et al. Antiretroviral drug resistance mutations sustain or enhance CTL recognition of common HIV-1 Pol epitopes. J Immunol 2004;172:7212–7219. doi: 10.4049/jimmunol.172.11.7212. Crossref, Medline, Google ScholarRussell RR, Bowmer MI, Nguyen C, et al. HIV-1 DNA burden in peripheral blood CD4+ cells influences disease progression, antiretroviral efficacy, and CD4+ T-cell restoration. Viral Immunol 2001;14:379–389. doi: 10.1089/08828240152716628. Link, Google ScholarFiguresReferencesRelatedDetails Volume 35Issue 5Jun 2022 InformationCopyright 2022, Mary Ann Liebert, Inc., publishersTo cite this article:Michael D. Grant and Rodney S. Russell.The Value of “00” One.Viral Immunology.Jun 2022.347-348.http://doi.org/10.1089/vim.2022.0087Published in Volume: 35 Issue 5: June 14, 2022PDF download

Following CARMENA and SURTIME, patients with metastatic renal cell carcinoma (mRCC) and International Metastatic RCC Database Consortium (IMDC) intermediate and poor risk receive systemic therapy with the primary tumour (primary) in place, with the option of deferred cytoreductive nephrectomy (CN) in responding patients. We retrospectively analysed the safety and efficacy of first-line nivolumab/ipilimumab in 71 primary mRCC patients (42.3% IMDC poor risk; 43.6% with more than three metastatic sites). The baseline mean primary diameter was 9.3 cm and median follow-up was 11.5 mo. Of 69 patients with at least one follow-up computed tomography scan, 23 (33.3 %) had a partial response (PR) of the primary after a median of 4.8 mo, which was associated with a 91.3% overall response rate at metastatic sites (MSs) and absence of progressive disease, irrespective of the IMDC risk. The complete response (CR) rate at MSs (n = 7 [10.1%]) is similar to the CR rate in CheckMate 214. Thirteen deferred CNs were performed (18.8%) after a median of 13 mo, rendering four patients disease free. Only 4.3% of primaries progressed; grade 3-4 immune-related adverse events occurred in 31.9%. Irrespective of the IMDC risk, patients with a PR in the primary had a 1-yr overall survival rate of 89% versus 67% in those without (p = 0.012). PATIENT SUMMARY:Patients with metastatic kidney cancer receiving immunotherapy with nivolumab and ipilimumab had superior response at metastatic sites and better survival irrespective of International Metastatic RCC Database Consortium (IMDC) risk.

Natural killer (NK) cells are important anti-viral effector cells. The function and phenotype of the NK cells that constitute an individual's NK cell repertoire can be influenced by ongoing or previous viral infections. Indeed, infection with human cytomegalovirus (HCMV) drives the expansion of a highly differentiated NK cell population characterized by expression of CD57 and the activating NKG2C receptor. This NK cell population has also been noted to occur in HIV-1-infected individuals. We evaluated the NK cells of HIV-1-infected and HIV-1-uninfected individuals to determine the relative frequency of highly differentiated CD57+NKG2C+ NK cells and characterize these cells for their receptor expression and responsiveness to diverse stimuli. Highly differentiated CD57+NKG2C+ NK cells occurred at higher frequencies in HCMV-infected donors relative to HCMV-uninfected donors and were dramatically expanded in HIV-1/HCMV co-infected donors. The expanded CD57+NKG2C+ NK cell population in HIV-1-infected donors remained stable following antiretroviral therapy. CD57+NKG2C+ NK cells derived from HIV-1-infected individuals were robustly activated by antibody-dependent stimuli that contained anti-HIV-1 antibodies or therapeutic anti-CD20 antibody, and these NK cells mediated cytolysis through NKG2C. Lastly, CD57+NKG2C+ NK cells from HIV-1-infected donors were characterized by reduced expression of the inhibitory NKG2A receptor. The abundance of highly functional CD57+NKG2C+ NK cells in HIV-1-infected individuals raises the possibility that these NK cells could play a role in HIV-1 pathogenesis or serve as effector cells for therapeutic/cure strategies.

Hybrid immunity induced by vaccination following recovery from SARS-CoV-2 infection is more robust than immunity induced by either infection or vaccination alone. To investigate how infection severity influenced the strength and character of subsequent vaccine-induced humoral or cellular immune responses against SARS-CoV-2, we assessed humoral and cellular immune responses against SARS-CoV-2 following recovery from infection, vaccine dose 1 and vaccine dose 2 in 35 persons recovered from COVID-19. Persons with polymerase chain reaction or serologically confirmed SARS-CoV-2 infection were recruited into a study of immunity against SARS-CoV-2. Self-reported symptoms categorized them as experiencing asymptomatic, mild, moderate or severe infection based on duration, intensity and need for hospitalization. Whole blood was obtained before vaccination and after first and second doses. Humoral immunity was assessed by ELISA and cellular immunity by ELISpot and intracellular flow cytometry. Responses were compared between groups recovered from either asymptomatic/mild (n = 14) or moderate/severe (n = 21) infection. Most subjects experienced robust increases in humoral and cellular immunity against SARS-CoV-2 spike (S) protein following 1 vaccination. Quantitative responses to second vaccination were marginal when measured 2.5 months afterwards and moderate or severe infection maintained stronger responses. Polyfunctional CD8+ T cell responses were largely restricted to subjects recovered from moderate or severe infection. One vaccine dose triggered stronger immune responses than in a comparable group never infected with SARS-CoV-2, while the second dose produced only minor lasting increases in humoral or cellular responses. Infection history should be considered in planning COVID-19 vaccine administration.

Since COVID-19 emerged in 2019, neutralizing antibody (Ab) responses have dominated as surrogate measures of protection against infection and severe illness. The mostly self-limiting course of SARS-CoV-2 infection suggests the innate immune system, including natural killer (NK) cells, plays a role in its control. Therefore, we investigated SARS-CoV-2-specific antibody-dependent cell-mediated cytotoxicity (ADCC) and if Abs against Wuhan and early SARS-CoV-2 variants activate ADCC against emergent highly transmissible variants. ADCC was evaluated by measuring NK cell cytotoxicity (51Cr) against plasma (1:1000)-sensitized MRC-5 cells stably transduced to express Wuhan or Delta spike. ADCC mediated by plasma from double-vaccinated and convalescent adults was generally low (range 1.2 – 24.9%; median 7.2% (IQR 5.2 – 10.6%), and plasma from those with previous natural infection mediated significant ADCC (> 10% lysis) to the same extent as plasma from fully vaccinated individuals (13/34 vs 11/44). Hybrid immunity imparted robust ADCC (27.9 ± 10.8% target lysis) in 32/34 individuals tested and levels of ADCC significantly correlated with IgG3 responses (P = 0.0093) against spike stalk (S2) domain. A peptide scan identified four distinct ‘hotspot’ regions associated with robust ADCC in individuals with reactive Abs. Overall, a minor decline in ADCC against Delta variant spike-expressing relative to Wuhan targets occurred in all groups; however, individuals with Abs against the region containing D614G mediated greater ADCC against Delta variant than Wuhan targets. Determining which Abs provide robust and broad ADCC can inform future COVID-19 prevention and treatment strategies and predict ADCC responses for emergent variants. Supported by grants from CIHR through the COVID Immunity Task Force (VR1-173202)

Reducing the socioeconomic toll from age-related physical and mental morbidities requires better understanding of factors affecting healthy aging. While many environmental, lifestyle, and genetic factors affect healthy aging, this study addressed the influence of cytomegalovirus (CMV) infection and immunity on age-related inflammation and cognitive abilities. Healthy adults 70–90 years old were recruited into a prospective study investigating relationships between anti-CMV immunity, markers of inflammation, baseline measures of cognitive ability, and changes in cognitive ability over 18 months. Humoral and cellular responses against CMV, levels of inflammatory markers, and cognitive abilities were measured at study entry, with measurement of cognitive abilities repeated 18 months later. CMV-seropositive and -seronegative sub-groups were compared, and relationships between anti-CMV immunity, markers of inflammation, and cognitive ability were assessed. Twenty-eight of 39 participants were CMV-seropositive, and two had CMV-specific CD8+ T cell responses indicative of CMV immune memory inflation. No significant differences for markers of inflammation or measures of cognitive ability were observed between groups, and cognitive scores changed little over 18 months. Significant correlations between markers of inflammation and cognitive scores with interconnection between anti-CMV antibody levels, fractalkine, cognitive ability, and depression scores suggest areas of focus for future studies.

4537 Background: Little is known regarding response and outcomes to ICI for patients (pts) with aUC who were previously treated with BCG for non-muscle invasive bladder cancer. We hypothesized that prior intravesical BCG would not be associated with changes in objective response or survival in pts with aUC treated with ICI. Methods: We performed a retrospective cohort study across 25 institutions. Demographic, intravesical BCG history, treatment and outcomes data were collected for pts with aUC who received ICI. Pts with aUC treated with ICIs were included, pts with pure non-UC, those treated with combination or on clinical trials, pts with multiple ICI treatment lines and those with upper tract UC were excluded. Pts were stratified to prior exposure versus no exposure to BCG. We compared overall response rate (ORR) using logistic regression; and progression-free survival (PFS) and overall survival (OS) using Kaplan-Meier and Cox proportional hazards. All analyses were performed in the overall population and further stratified by treatment line (first-line [1L] vs salvage [2+L]) and multivariable models. The stratified analysis was also adjusted for an internally developed risk score for 1L and Bellmunt risk score for 2+L; p<0.05 was significant. Results: 1026 aUC pts treated with ICI were identified; 614 pts, 617 pts, and 641 pts were included in ORR, OS and PFS analyses, respectively. Overall, mean age at CPI initiation was 70, 76% were men, 70% were current or former smokers, 75% White, 29% with mixed histology, and 24% had prior exposure to BCG. ORR to ICI in pts with or without prior exposure to BCG was similar, 27% and 28% respectively (OR=0.93 [95% CI 0.61-1.42], p=0.73). Median OS (mOS) for pts with vs without prior BCG exposure was 9 vs 10 mo (HR=1.13 [95% CI 0.88-1.44], p=0.35). Median PFS (mPFS) was 4 months (mo) in both groups (HR=1.02 [95% CI 0.82-1.27], p=0.83). ORR, PFS and OS analyses stratified by ICI treatment line (1L vs 2+L) are listed in the table. Conclusions: In this multi-institutional retrospective analysis, prior intravesical BCG was not associated with objective response or survival in pts with aUC treated with ICI. Limitations of this study include retrospective nature, lack of randomization and possible confounding, but it does provide important preliminary data that selection for ICI treatment should not be impacted by prior exposure to BCG. Further clinical and molecular biomarker exploration is needed to refine patient selection for ICI in aUC.[Table: see text]