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Introduction: Human peripheral blood mononuclear cells (hPBMCs) are widely used in fundamental research and clinical applications as studying their responses to in vitro activation is an effective way to uncover functional alterations and disease associated phenotypes. However, the availability of samples in large numbers at a specific time and location remains challenging, hence they often might preferably be collected and cryopreserved for later analysis. While the effect of cryopreservation on viability and cell surface expression is well established, changes in activity and cytokine secretion still lead to conflicting results as it is often measured in bulk or within the cells. Methods: Here, we used our platform for dynamic single-cell multiplexed cytokine secretion measurement and compared it to a traditional intracellular cytokine staining to quantify the effect of cryopreservation on cytokine secretion and expression of individual hPBMCs. Results: Following stimulation with LPS or anti-CD3/CD28 antibodies for up to 36 or 72 h incubation, we observed distinct alterations in cytokine responses due to cryopreservation when comparing to fresh samples, but also remarkable consistencies for some cytokines and parameters. In short, the frequencies of cytokine-secreting cells in cryopreserved samples were lower for IL-6 (LPS), IL1-b (CD3/CD28) and IFN-g (CD3/CD28), while the frequency and dynamics of IL-8 secretion were strongly impacted in all cases. We observed a large disconnect between cytokine expression and secretion for TNF-a, where the expression dramatically increased after cryopreservation, but actual secretion was, in comparison, remarkably stable. The polyfunctionality of single cells was altered by cryopreservation in specific co-secreting populations led by the effects on IL-6 or IL-8 secretion. Among immune cells, cryopreservation seemed to affect lymphocytes and monocytes differently as effects appeared early on in lymphocytes while generally observed in later time points in monocytes. Conclusion: Together, this study offers an in-depth quantitative insight into the biological behavior of immune cells in response to cryopreservation and stimulation, further providing some insights into conflicting results in the literature as well as guidelines for researchers planning to assess cytokine-secreting from frozen hPBMCs in immunological research or clinical applications. [ABSTRACT FROM AUTHOR]

Cytomegalovirus (CMV)-seropositive adults have large T cell responses to a wide range of CMV proteins; these responses have been associated with chronic inflammation and frailty in people with or without HIV infection. We analyzed the relationships between chronic HIV infection, frailty, and the breadth and polyfunctionality of CD4 and CD8 T cell responses to CMV. Peripheral blood mononuclear cells from 42 men (20 without HIV and 22 with virologically suppressed HIV) in the Multicenter AIDS Cohort Study (MACS) were stimulated with peptide pools spanning 19 CMV open reading frames (ORFs). As measured by flow cytometry and intracellular cytokine staining for IFN-γ, TNF-α, and IL-2, CD8 T cells from men with HIV responded to significant ly more CMV ORFs than those from men without HIV. This was primarily due to a broader response to ORFs that are expressed during the late phase of CMV replication. The number of ORFs to which a participant's T cells responded was positively correlated with the sum of all that individual's T cell responses; these correlations were weaker in men with than without HIV. Polyfunctional CMV-specific CD4 responses (production of more than one cytokine) were significantly lower in men with than without HIV. Frailty status did not substantially affect the breadth or magnitude of the CMV-specific T cell responses. These results suggest that immune control of CMV infection is affected more by chronic HIV infection than by frailty. The differences between men with and without HIV were similar to those reported between young and older adults without HIV. [ABSTRACT FROM AUTHOR]

Words expressing 'make' are semantically and functionally versatile in many languages. Focusing on the word makee , derived from English make , in Chinese Pidgin English, a contact language originated in 18th-century Canton, China, this paper examines its origins and functions. Primarily a lexical verb, makee is polyfunctional in that it also occurs in the complex form [ makee X] which indicates causation, imperative, and verbalization. Functioning as a main verb and a causative verb, makee largely mirrors the corresponding words in the source languages of the pidgin, namely English and Cantonese. However, makee as an imperative marker and a verbalizer are innovative features. In imperative sentences, makee in combination with another verb indicate commands. As a verbalizer, makee gives a verbal interpretation to the whole construction. These two new features of makee are not attributable to existing models in the source languages. [ABSTRACT FROM AUTHOR]

Several studies have investigated the antibody response to SARS-CoV-2, focusing particularly on the systemic humoral immune response and the production of immunoglobulin G (IgG) antibodies. IgA antibodies play a crucial role in protecting against respiratory viral infections but have also been associated with the pathophysiology of COVID-19. We performed a prospective study of 169 COVID-19 patients – 50 with critical/severe (ICU), 47 with moderate (Non-ICU), and 72 with asymptomatic COVID-19 – to explore the humoral immune response to SARS-CoV-2 infection. We found that the early systemic IgA response strongly induced in patients with severe disease did not block IgG neutralization functions and activated FcRs more effectively than IgG. However, even if SIgA levels were high, mucosal IgA antibodies could not control the infection effectively in patients with severe disease. Our findings highlight the complexity of the immune response to SARS-CoV-2 exhibiting high systemic levels of IgA with strong neutralizing capacity in severe cases, together with higher levels of IgA-FcR activation than in asymptomatic patients. They also suggest the need for further research to fully understand the role of IgA and its structural alterations in mucosal tissues in cases of severe disease and the impact of these antibodies on disease progression.

Reportative evidential markers are – in contrast to other evidential markers – compatible with distancing interpretations, in which the speaker denies the truth of what is being reported. This exceptional behaviour of reportatives is termed 'reportative exceptionality' (AnderBois 2014). In this paper, which addresses French, Dutch and German reportative markers, we argue that they differ with respect to the frequency with which such distancing interpretations actually arise. The French reportative conditionnel most frequently occurs with distancing interpretations, whereas German sollen hardly occurs with this function. Dutch zou takes up an intermediate position. It is claimed that the higher compatibility of the conditionnel with distancing interpretations can be accounted for by a number of factors: its general preference for contexts in which other perspectives than the speaker's are highly salient; the fact that it has past tense morphology; and its general semantic make-up in which the marking of hypotheticality is a key function. [ABSTRACT FROM AUTHOR]

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The linguistic contact between Slovak and French gave birth to more than 3,000 loan, simple and complex words. Of the 748 simple words, nouns account for a major proportion of 642 lexemes. The present study typologizes, systematizes and analyses the process of their morphological adaptation, preceded by more or less significant change of pronunciation, orthography and grammatical gender assignment, in the transition from a predominantly analytic language (French) to a flectional language (Slovak). In certain cases, the inflectional formants of noun Gallicisms in Slovak carry the French derivational formants, they are characterized by a certain degree of polyfunctionality and require a more detailed qualitative and quantitative analysis which shows that, occasionally, in the case of French derivatives, the Slovak Gallicisms carry over even the morphological (sometimes also onomasiological) structure from Latin.

IntroductionHuman peripheral blood mononuclear cells (hPBMCs) are widely used in fundamental research and clinical applications as studying their responses to in vitro activation is an effective way to uncover functional alterations and disease associated phenotypes. However, the availability of samples in large numbers at a specific time and location remains challenging, hence they often might preferably be collected and cryopreserved for later analysis. While the effect of cryopreservation on viability and cell surface expression is well established, changes in activity and cytokine secretion still lead to conflicting results as it is often measured in bulk or within the cells.MethodsHere, we used our platform for dynamic single-cell multiplexed cytokine secretion measurement and compared it to a traditional intracellular cytokine staining to quantify the effect of cryopreservation on cytokine secretion and expression of individual hPBMCs.ResultsFollowing stimulation with LPS or anti-CD3/CD28 antibodies for up to 36 or 72 h incubation, we observed distinct alterations in cytokine responses due to cryopreservation when comparing to fresh samples, but also remarkable consistencies for some cytokines and parameters. In short, the frequencies of cytokine-secreting cells in cryopreserved samples were lower for IL-6 (LPS), IL1-β (CD3/CD28) and IFN-γ (CD3/CD28), while the frequency and dynamics of IL-8 secretion were strongly impacted in all cases. We observed a large disconnect between cytokine expression and secretion for TNF-α, where the expression dramatically increased after cryopreservation, but actual secretion was, in comparison, remarkably stable. The polyfunctionality of single cells was altered by cryopreservation in specific co-secreting populations led by the effects on IL-6 or IL-8 secretion. Among immune cells, cryopreservation seemed to affect lymphocytes and monocytes differently as effects appeared early on in lymphocytes while generally observed in later time points in monocytes.ConclusionTogether, this study offers an in-depth quantitative insight into the biological behavior of immune cells in response to cryopreservation and stimulation, further providing some insights into conflicting results in the literature as well as guidelines for researchers planning to assess cytokine-secreting from frozen hPBMCs in immunological research or clinical applications.

Previous work on grammaticalization in Arabic has focused on the development of a number of morphosyntactic features such as mood, tense and aspect, and to a lesser degree modal features and discourse features in verbs, prepositions and conjunctions in the modern Arabic vernaculars or in Standard Arabic. Using grammaticalization theory, this study demonstrates with diachronic data from Classical Arabic and synchronic data from Iraqi Arabic that some Classical Arabic nouns have evolved into Iraqi Arabic degree adverbs, and in some cases into indefinite quantifiers as well. The grammaticalizated terms exhibit the characteristic outcomes of grammaticalization, namely semantic extension or bleaching, decategorization, and phonetic erosion. Reanalysis is shown to have played a decisive role in the development of the grammaticalized terms. Specific contexts of use of the Classical Arabic nouns are shown to have triggered the grammaticalization process. The proposed grammaticalization is shown to have been driven by two Minimalist economy principles. Data from neighboring Arabic vernaculars seem to validate the proposed analysis. The suggestion is made that the functional versatility of CA nouns might have contributed to their being the target of the proposed grammaticalization. [ABSTRACT FROM AUTHOR]

Cytokine polyfunctionality is a well-established concept in immune cells, especially T cells, and their ability to concurrently produce multiple cytokines has been associated with better immunological disease control and subsequent effectiveness during infection and disease. To date, only little is known about the secretion dynamics of those cells, masked by the widespread deployment of mainly time-integrated endpoint measurement techniques that do not easily differentiate between concurrent and sequential secretion. Here, we employed a single-cell microfluidic platform capable of resolving the secretion dynamics of individual PBMCs. To study the dynamics of poly-cytokine secretion, as well as the dynamics of concurrent and sequential polyfunctionality, we analyzed the response at different time points after ex vivo activation. First, we observed the simultaneous secretion of cytokines over the measurement time for most stimulants in a subpopulation of cells only. Second, polyfunctionality generally decreased with prolonged stimulation times and revealed no correlation with the concentration of secreted cytokines in response to stimulation. However, we observed a general trend towards higher cytokine secretion in polyfunctional cells, with their secretion dynamics being distinctly different from mono-cytokine-secreting cells. This study provided insights into the distinct secretion behavior of heterogenous cell populations after stimulation with well-described agents and such a system could provide a better understanding of various immune dynamics in therapy and disease.

The research aims to conduct a functional analysis of the opposing conceptual metaphors ‘heaven’ and ‘hell’ within English headlines from BBC and The Guardian news between 1999 and 2023. The relevance of this study lies in the need to explore the lexical meaning of fundamental theological terms based on the unreal cognitive constructs in the context of evolving social paradigms. Various research methods were employed, including continuous sampling, classification, thematic analysis, definition analysis, and semantic analysis.The findings indicate that the conceptual metaphor ‘hell’ possesses a broader semantic field compared to ‘heaven,’ both in terms of definition and function. Metaphorical usage is prevalent, but headlines also feature ‘heaven’ and ‘hell’ within idiomatic, emphatic, allusive, direct, zeugmatic phrases, and contaminations. These metaphors encompass thematic areas with polar connotational meanings:Hell: Politics (political instability, struggle, political figure replacement, anti-crisis measures), Social life (reforms, discrimination, limitations, mandatory expenses), Crimes (abuses, threats, harassment, assaults, victims), Nature (natural disasters, difficult conditions, environmental catastrophes), Sport (injuries, competition, psychological attacks, overcoming), Entertainment (poor quality service, bad habits, imbalance, disruptions), Travel (unforeseen circumstances, transport collapses), Culture (culture shock).Heaven: Politics (political decisions, electoral victories), Social life (festivals, feasts, family relations, relationships between generations), Crimes (controversy resolution, search for solutions), Nature (biodiversity, renewal of nature), Sport (victories, lucrative contracts, goal achievement), Entertainment (parties, bright performances, romanticism), Travel (pleasure, rest, nature), Culture (acquaintance with outstanding cultural objects, visiting cultural events).This research demonstrates that the dichotomous representation of the conceptual metaphors ‘heaven’ and ‘hell’ is marked by polyfunctionality, driven by the universal evaluative component inherent in their meaning, rooted in the implied polarization of the concepts ‘good’ and ‘bad.’ The emotional and moral dimensions contribute to heightened news perception and effective communication within media discourse.

The Hebrew negation adverbial bilti 'not' seems to function very differently in Biblical Hebrew than it does in Contemporary Hebrew. This paper addresses this difference and discusses its evolution. The main question addressed in this paper is: How has Hebrew bilti, originally an exceptive marker (with sentential scoping), ended up functioning solely as a privative in contemporary Hebrew? First, this paper argues that the biblical usage of bilti was expanded and turned into a polyfunctional (or 'polysemous') item. This happened via a constructionalization process which led to grammatical changes ('grammaticalization'): The initially implicated negation (via a generalized implicature) turned explicit (semantic). In addition, in Hebrew's later periods, the usage of bilti was narrowed and it became a privative. Thus, firstly, a pragmatically motivated path of constructionalization of bilti in Biblical Hebrew is suggested. That is, the "pragmatic negation" that arose via a generalized implicature shifted to the semantic level (performing semantic negation, explicit negation). Secondly, bilti's functions in post-biblical Hebrew periods are outlined, tracing its narrowing functions until its fixation in Contemporary Hebrew as a privative. [ABSTRACT FROM AUTHOR]

Copyright of Studies about Language / Kalbu Studijos is the property of Studies about Language / Kalbu Studijos and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Immunocompromised individuals, particularly autologous hematopoietic stem cell transplant (auHSCT) recipients, are at high risk for herpes zoster (HZ). We provide an in-depth description of humoral and cell-mediated immune (CMI) responses by age (protocol-defined) or underlying disease (post-hoc) as well as efficacy by underlying disease (post-hoc) of the adjuvanted recombinant zoster vaccine (RZV) in a randomized observer-blind phase III trial (ZOE-HSCT, NCT01610414). 1846 adult auHSCT recipients were randomized to receive a first dose of either RZV or placebo 50-70 days post-auHSCT, followed by the second dose at 1-2 months (M) later. In cohorts of 114-1721 participants, at 1 M post-second vaccine dose: Anti-gE antibody geometric mean concentrations (GMCs) and median gE-specific CD4[2+] T-cell frequencies (CD4 T cells expressing ≥2 of four assessed activation markers) were similar between 18-49 and ≥50-year-olds. Despite lower anti-gE antibody GMCs in non-Hodgkin B-cell lymphoma (NHBCL) patients, CD4[2+] T-cell frequencies were similar between NHBCL and other underlying diseases. The proportion of polyfunctional CD4 T cells increased over time, accounting for 79.6% of gE-specific CD4 T cells at 24 M post-dose two. Vaccine efficacy against HZ ranged between 42.5% and 82.5% across underlying diseases and was statistically significant in NHBCL and multiple myeloma patients. In conclusion, two RZV doses administered early post-auHSCT induced robust, persistent, and polyfunctional gE-specific immune responses. Efficacy against HZ was also high in NHBCL patients despite the lower humoral response.

Background Despite wide use of adjuvanted influenza vaccine in nursing home residents (NHR), little immunogenicity data exist for this population. Methods We collected blood from NHR (n = 85) living in nursing homes participating in a cluster randomized clinical trial comparing MF59-adjuvanted trivalent inactivated influenza vaccine (aTIV) with nonadjuvanted vaccine (TIV) (parent trial, NCT02882100). NHR received either vaccine during the 2016–2017 influenza season. We assessed cellular and humoral immunity using flow cytometry and hemagglutinin inhibition, antineuraminidase (enzyme-linked lectin assay), and microneutralization assays. Results Both vaccines were similarly immunogenic and induced antigen-specific antibodies and T cells, but aTIV specifically induced significantly larger 28 days after vaccination (D28) titers against A/H3N2 neuraminidase than TIV. Conclusions NHRs respond immunologically to TIV and aTIV. From these data, the larger aTIV-induced antineuraminidase response at D28 may help explain the increased clinical protection observed in the parent clinical trial for aTIV over TIV in NHR during the A/H3N2-dominant 2016–2017 influenza season. Additionally, a decline back to prevaccination titers at 6 months after vaccination emphasizes the importance of annual vaccination against influenza. Clinical Trials Registration NCT02882100. [ABSTRACT FROM AUTHOR]

Acute T-cell-mediated rejection (aTCMR) still remains a clinical problem after kidney transplantation despite significant improvements in immunosuppressive regimens. Polyfunctional T cells, i.e. T cells producing multiple pro-inflammatory cytokines, are believed to be the most relevant T cells in an immune response. The aim of this study was to determine whether polyfunctional donor-reactive T cells are associated with aTCMR. In a case–control study, 49 kidney transplant recipients with a biopsy-proven aTCMR in the first year after transplantation were included, as well as 51 controls without aTCMR. Circulating donor-reactive T cells were identified by the expression of CD137 after short-term co-culture with donor antigen-presenting cells. Polyfunctional donor-reactive T cells were further characterized by dissection into different T-cell subsets encompassing the spectrum of naïve to terminally differentiated effector T cells. Prior to kidney transplantation, proportions of donor-reactive CD4+ (0.03% versus 0.02%; P < 0.01) and CD8+ (0.18% versus 0.10%; P < 0.01) CD137++ T cells were significantly higher in recipients with a biopsy-proven aTCMR versus non-rejectors. Polyfunctionality was higher (P = 0.03) in this subset of CD137-expressing T cells. These cells were predominantly of the EM/EMRA-phenotype, with polyfunctional donor-reactive CD137++CD4+ T cells predominantly co-expressing CD28 whereas approximately half of the polyfunctional CD137++CD8+ T cells co-expressed CD28. In addition, at the time of aTCMR, polyfunctional donor-reactive CD137++ CD4+, but not CD8+, T cells, were specifically decreased by 75% compared to before transplantation in recipients with as well as those without an aTCMR. Prior to transplantation, the proportion of polyfunctional donor-reactive CD137++ T cells is associated with the occurrence of a biopsy-proven aTCMR within the first year after transplantation. Acute TCMR remains a clinical problem after kidney transplantation despite significant improvements in immunosuppressive regimens. Poly-functional (producing ≥2 cytokines) T cells are believed to be the most relevant T cells in an immune response. Prior to kidney transplantation proportions of poly-functional CD137 high expressing donor-reactive CD4+ and CD8+ T cells are higher in recipients experiencing an acute TCMR within the first year after transplantation compared to those who remained free of rejection. [ABSTRACT FROM AUTHOR]

Discourse markers (DMs) are generally described as polyfunctional. There are different approaches to polyfunctionality (homonymy, monosemy, polysemy), which have been widely explored theoretically; however, there are not many experimental approaches showing how polyfunctionality works (i.e., how it is assimilated by speakers or readers when it emerges in various real communicative contexts). This paper presents results on experimental polyfunctionality in DMs through eye-tracking methods applied to the Spanish marker o sea ‘I mean’. The main question addressed is how polyfunctionality is understood in terms of ocular behavior: specifically, if processing results reveal a monosemic or a polysemic pattern for this marker.

Polyfunctionality refers to cases in which the same formal material is systematically reused with different functions. It represents a type of complexity of exponence, wherein there is a non-one-to-one mapping between function and form. Much recent work in morphology has emphasized the role of implicative structure in resolving the communicative challenges associated with complex form-function mappings. However, previous work has focused almost entirely on complexity of exponence as a challenge for the speaker predicting novel forms (knowing how to encode information), and very little on the challenge for the listener in decoding novel forms. It also has focused almost exclusively on inflectional paradigms, and has not explored how other types of implicative structure might be important in languages with diverse morphological systems. This paper investigates the role of syntagmatic (i.e. word-internal) implicative structure in Ket (Yeniseian, Siberia), a polysynthetic language with numerous polyfunctional markers. It is shown that such markers are organized into networks of implicative relations with one another, wherein less polyfunctional markers along a scale of polyfunctionality disambiguate the function of more polyfunctional markers. This allows uncertainty with regard to their function in any particular instance to remain low. The Ket data make wider typological predictions for the relationship between complexity of exponence and implicative structure. [ABSTRACT FROM AUTHOR]

This paper addresses a discourse particle that typically appears in conceptually oral / colloquial interaction and instantiates one of many different functions of the lexical element so (lit. ‘so, this way’) in Present-Day German. In contrast to the modal adverb so and just as some of its other polysemous counterparts, this element appears to be utterly desemanticized and non-referential, thereby modifying the meaning of the sentence at a very abstract level. On the basis of its distributional properties, its semantic contribution to the host utterance, as well as of independent assumptions on the notion of modality, it is contended that this so is to be categorized as an interrogative modal particle whose primary role lies in managing the Common Ground in non-monologic contexts such that the speaker requires the addressee to fill a gap in their knowledge of a given situation with the additional implication that the relevant piece of information does not have to be exhaustive. The aim of this paper is not to provide a comprehensive analysis of the phenomenon, but rather to make a theoretical proposal regarding the status of this element and thereby stimulate the discussion on the defining features of modal particles. [ABSTRACT FROM AUTHOR]

This article investigates the semantic polyfunctionality of affixes, i.e. their ability to serve a variety of distinct semantic functions. Based on the analysis of a sample of 3,091 deverbal nouns ending with 46 different suffixes in French, the study examines the diversity of semantic functions realized by deverbal suffixes, the distribution of these functions across suffixes and the relationships that may exist between different functions. It appears that polyfunctionality is widespread among French deverbal suffixes and involves a large number of semantic functions, with highly variable realization frequency. Several fundamental aspects of affix polyfunctionality are further highlighted. A probabilistic analysis shows that polyfunctionality can be driven by non-arbitrary semantic associations between functions. A hierarchy of functions can also be postulated depending on whether they can be instantiated by monosemous or only polysemous derivatives. In addition, polyfunctionality appears to be inseparable from rivalry relationships and to determine the degree of rivalry between affixes. Overall, the study illustrates that affix polyfunctionality is governed by systematic organizing principles whose ramifications touch on lexical ambiguity and morphological competition. [ABSTRACT FROM AUTHOR]

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The German proverb “Es ist dafür gesorgt, daß die Bäume nicht in den Himmel wachsen“ with its shortened variant „Die Bäume wachsen nicht in den Himmel“ has been transmitted since the early sixteenth century. Its written documentation begins 1526 with Martin Luther, and it appears since 1590 in numerous variants in proverb collections. Goethe quoted it in his autobiography, and it is present in the works of Heinrich Heine, Joseph von Eichendorff, Georg Herwegh, Gottfried Keller, Theodor Fontane, Wilhelm Raabe, Hermann Hesse, Alfred Andersch, and others. Max Weber and Rosa Luxemburg made socio-political use of it, and that is also true for Winston S. Churchill, who played a part in distributing it in English translation as „Care is taken that trees don’t grow to the sky“ and „Trees don’t grow to the sky“. Joseph Goebbels quotes it repeatedly as a propagandistic leitmotif, and it also plays a role in political contexts by chancellors Conrad Adenauer, Willy Brandt, and Helmut Schmidt. Especially aphoristic writers as Dietmar Beetz, Erwin Chargaff, Peter Maiwald, Felix Renner, and Gerhard Uhlenbruck have dealt with it critically by changing it into anti-proverbs. By way of many contextualized references it is shown how the proverb developed during five centuries and how it is marked to this day by its polysituativity, polyfunctionality, and polysemanticity.

In line with several authors (just to give an example, Aikhenvald (2015) who argue that the intersection of different inflective categories is the key to the "polyfunctionality" characteristic of verb forms, we examine the confluence of a certain range of temporal and aspectual information about modal auxiliaries in the determination of specific behaviors. As in current Engl. except for will (

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Background: Predictive biomarkers for immune checkpoint inhibitors (ICIs), e.g., programmed death ligand-1 (PD-L1) tumor proportional score (TPS), remain limited in clinical applications. Predictive biomarkers that require invasive tumor biopsy procedures are practically challenging especially when longitudinal follow-up is required. Clinical utility of tissue-based PD-L1 TPS also becomes diluted when ICI is combined with chemotherapies. Peripheral single T-cell dynamic polyfunctionality profiling offers the opportunity to reveal rare T-cell subpopulations that are polyfunctional and responsible for the underlying ICI treatment molecular response that bulk biological assays cannot achieve. Here, we evaluated a novel live single-cell functional liquid biopsy cytokine profiling platform, IsoLight, as a potential predictive biomarker to track ICI treatment response and clinical outcomes in non-small cell lung cancer (NSCLC)., Methods: Peripheral blood mononuclear cell samples of 10 healthy donors and 10 NSCLC patients undergoing ICI-based therapies were collected longitudinally pre-/post-ICI treatment after ≥2 cycles under institutional review board (IRB)-approved protocols. Cancer blood samples were collected from unresectable advanced stage (III-IV) NSCLC patients. Clinical course and treatment response and survival outcomes were extracted from electronic health records, with treatment response assessed by treating oncologists based on RECIST. CD4 + and CD8 + T-cells were enriched magnetically and analyzed on the IsoLight platform. Single T-cells were captured in microchambers on IsoCode chips for proteomic immune cytokines profiling. Functional polyfunctionality data from 55,775 single cells were analyzed with IsoSpeak software, 2D- and 3D-t-distributed stochastic neighbor embedding (t-SNE) analysis, kappa coefficient, and Kaplan-Meier survival plots. P values ≤0.05 is considered statistically significant., Results: Pre-treatment baseline polyfunctionality profiles could not differentiate NSCLC patients from healthy subjects, and could not differentiate ICI responders from non-responders. We found a statistically significant difference between responders and non-responders in CD8 + T-cells' changes in overall polyfunctionality (ΔPolyFx) (P=0.01) and polyfunctional strength index (ΔPSI) (P=0.006) in our dynamic pre-/post-treatment single cell measurements, both performing better than PD-L1 TPS alone (P=0.08). In the 3D-t-SNE analysis, subpopulations of post-treatment CD8 + T-cells in ICI responders displayed distinct immune cytokine profiles from those in pre-treatment cells. CD8 + T-cells ΔPolyFx and ΔPSI scores performed better than PD-L1 TPS in ICI response correlation. Moreover, combined PD-L1 strong TPS and ΔPSI >15 scores strongly correlated with early ICI response with a robust kappa coefficient of 1.0 (P=0.003), which was previously statistically established to indicate a perfect agreement between the prediction and actual response status. Interestingly, high CD4 + T-cells ΔPSI >5 was found to correlate with a strong trend of improved progression-free survival (3.9-fold) (10.8 vs. 2.8 months; P=0.07) and overall survival (3-fold) (34.5 vs. 11.5 months; P=0.09) in ICI-treated patients., Conclusions: Our study nominates single peripheral T-cell polyfunctionality dynamics analysis to be a promising liquid biopsy platform to determine potential ICI predictive biomarker in NSCLC. It warrants further studies in larger prospective cohorts to validate the clinical utilities and to further optimize cancer immunotherapy., Competing Interests: Conflicts of Interest: All authors have completed the ICMJE uniform disclosure form (available at https://tlcr.amegroups.com/article/view/10.21037/tlcr-24-260/coif). P.C.M. received consulting honorarium from AstraZeneca, BeiGene, Pfizer and Daiichi Sankyo; and conference traveling support from DAVA Oncology; and clinical trial research grants to the institution from Apollomics, BeiGene, AstraZeneca, OncoC4, GenMab, Elevation Oncology, Merck, AbbVie and Mirati. The other authors have no conflicts of interest to declare., (2024 AME Publishing Company. All rights reserved.)

The formation of a robust long-term antigen (Ag)-specific memory, both humoral and cell-mediated, is created following severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection or vaccination. Here, by using polychromatic flow cytometry and complex data analyses, we deeply investigated the magnitude, phenotype, and functionality of SARS-CoV-2-specific immune memory in two groups of healthy subjects after heterologous vaccination compared to a group of subjects who recovered from SARS-CoV-2 infection. We find that coronavirus disease 2019 (COVID-19) recovered patients show different long-term immunological profiles compared to those of donors who had been vaccinated with three doses. Vaccinated individuals display a skewed T helper (Th)1 Ag-specific T cell polarization and a higher percentage of Ag-specific and activated memory B cells expressing immunoglobulin (Ig)G compared to those of patients who recovered from severe COVID-19. Different polyfunctional properties characterize the two groups: recovered individuals show higher percentages of CD4+ T cells producing one or two cytokines simultaneously, while the vaccinated are distinguished by highly polyfunctional populations able to release four molecules, namely, CD107a, interferon (IFN)-γ, tumor necrosis factor (TNF), and interleukin (IL)-2. These data suggest that functional and phenotypic properties of SARS-CoV-2 adaptive immunity differ in recovered COVID-19 individuals and vaccinated ones.

Epoxidation of linseed oil fatty acid ethyl esters promoted by a series of functionalized imidazolium ionic liquids was studied. In this reaction acidic imidazolium ILs behave not only as an acid catalysts but also play an additional role as a specific activator for the reaction of epoxidation of unsaturated fatty derivatives in H2O2/formic acid oxidation system. The obtained results indicate that the hydrogen‐bonding interaction of imidazolium cation with epoxide group influences the selectivity of the epoxidation reaction. In particular, the presence of unprotected weak "acid" C2‐H proton significantly lowers the selectivity to hydroxyl derivatives. The obtained results also point to another example of specific polyfunctionality of ILs, especially imidazolium one, used as surfactants and process activators. [ABSTRACT FROM AUTHOR]

Immunocompromised individuals, particularly autologous hematopoietic stem cell transplant (auHSCT) recipients, are at high risk for herpes zoster (HZ). We provide an in-depth description of humoral and cell-mediated immune (CMI) responses by age (protocol-defined) or underlying disease (post-hoc) as well as efficacy by underlying disease (post-hoc) of the adjuvanted recombinant zoster vaccine (RZV) in a randomized observer-blind phase III trial (ZOE-HSCT, NCT01610414). 1846 adult auHSCT recipients were randomized to receive a first dose of either RZV or placebo 50–70 days post-auHSCT, followed by the second dose at 1–2 months (M) later. In cohorts of 114–1721 participants, at 1 M post-second vaccine dose: Anti-gE antibody geometric mean concentrations (GMCs) and median gE-specific CD4[2+] T-cell frequencies (CD4 T cells expressing ≥2 of four assessed activation markers) were similar between 18–49 and ≥50-year-olds. Despite lower anti-gE antibody GMCs in non-Hodgkin B-cell lymphoma (NHBCL) patients, CD4[2+] T-cell frequencies were similar between NHBCL and other underlying diseases. The proportion of polyfunctional CD4 T cells increased over time, accounting for 79.6% of gE-specific CD4 T cells at 24 M post-dose two. Vaccine efficacy against HZ ranged between 42.5% and 82.5% across underlying diseases and was statistically significant in NHBCL and multiple myeloma patients. In conclusion, two RZV doses administered early post-auHSCT induced robust, persistent, and polyfunctional gE-specific immune responses. Efficacy against HZ was also high in NHBCL patients despite the lower humoral response.

Selective immune pressure contributes to relapse due to target antigen downregulation in patients treated with anti-CD19 chimeric antigen receptor (CAR) T cells. Bispecific lentiviral anti-CD20/anti-CD19 (LV20.19) CAR T cells may prevent progression/relapse due to antigen escape. Highly polyfunctional T cells within a CAR T-cell product have been associated with response in single-antigen-targeted anti-CD19 CAR T cells. The authors performed a single-cell proteomic analysis to assess polyfunctional cells in our LV20.19 CAR T-cell product. Analysis was limited to those treated at a fixed dose of 2.5 × 106 cells/kg (n = 16). Unused pre-infusion CAR T cells were thawed, sorted into CD4/CD8 subsets and stimulated with K562 cells transduced to express CD19 or CD20. Single-cell production of 32 individual analytes was measured and polyfunctionality and polyfunctional strength index (PSI) were calculated. Fifteen patients had adequate leftover cells for analysis upon stimulation with CD19, and nine patients had adequate leftover cells for analysis upon stimulation with CD20. For LV20.19 CAR T cells, PSI was 866–1109 and polyfunctionality was 40–45%, which were higher than previously reported values for other CAR T-cell products. Stimulation with either CD19 or CD20 antigens resulted in similar levels of analyte activation, suggesting that this product may have efficacy in CD19– patient populations. [ABSTRACT FROM AUTHOR]

Specific T cells are crucial to control SARS-CoV-2 infection, avoid reinfection and confer protection after vaccination. We have studied patients with severe or moderate COVID-19 pneumonia, compared to patients who recovered from a severe or moderate infection that had occurred about 4 months before the analyses. In all these subjects, we assessed the polyfunctionality of virus-specific CD4+ and CD8+ T cells by quantifying cytokine production after in vitro stimulation with different SARS-CoV-2 peptide pools covering different proteins (M, N and S). In particular, we quantified the percentage of CD4+ and CD8+ T cells simultaneously producing interferon-γ, tumor necrosis factor, interleukin (IL)-2, IL-17, granzyme B, and expressing CD107a. Recovered patients who experienced a severe disease display high proportions of antigen-specific CD4+ T cells producing Th1 and Th17 cytokines and are characterized by polyfunctional SARS-CoV-2-specific CD4+ T cells. A similar profile was found in patients experiencing a moderate form of COVID-19 pneumonia. No main differences in polyfunctionality were observed among the CD8+ T cell compartments, even if the proportion of responding cells was higher during the infection. The identification of those functional cell subsets that might influence protection can thus help in better understanding the complexity of immune response to SARS-CoV-2. [ABSTRACT FROM AUTHOR]

Copyright of CIRCULO de Linguistica Aplicada a la Comunicacion is the property of Universidad Complutense de Madrid and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Discourse markers (DMs) are generally described as polyfunctional. There are different approaches to polyfunctionality (homonymy, monosemy, polysemy), which have been widely explored theoretically; however, there are not many experimental approaches showing how polyfunctionality works (i.e., how it is assimilated by speakers or readers when it emerges in various real communicative contexts). This paper presents results on experimental polyfunctionality in DMs through eye-tracking methods applied to the Spanish marker o sea ‘I mean’. The main question addressed is how polyfunctionality is understood in terms of ocular behavior: specifically, if processing results reveal a monosemic or a polysemic pattern for this marker. [ABSTRACT FROM AUTHOR]

Association of HIV-1-specific T-cell responses to infection risk in seronegative individuals is controversial. We quantified and phenotypically characterized gp120-specific T-cell responses in HIV-1 exposed, but uninfected subjects enrolled in the global Pre-exposure Prophylaxis Initiative (iPrEx) chemoprophylaxis trial. IFNγ ELISpot responses were detected in 24% of subjects irrespective of infection outcome. HIV-1 gp120 envelope-specific T-cell responses were more uniformly IFN-γ+TNF-α+Mip-1β+ in persistently seronegative subjects relative to subjects who later seroconverted (median frequency of 76.5% and 66.5%, respectively). IFNγ responses targeted the V2 loop for subjects who remained seronegative. HIV-1 gp120 envelope V2 loop-specific CD8 T-cell responses may help to protect against HIV-1 acquisition.

Association of HIV-1-specific T-cell responses to infection risk in seronegative individuals is controversial. We quantified and phenotypically characterized gp120-specific T-cell responses in HIV-1 exposed, but uninfected subjects enrolled in the global Pre-exposure Prophylaxis Initiative (iPrEx) chemoprophylaxis trial. IFNγ ELISpot responses were detected in 24% of subjects irrespective of infection outcome. HIV-1 gp120 envelope-specific T-cell responses were more uniformly IFN-γ+TNF-α+Mip-1β+ in persistently seronegative subjects relative to subjects who later seroconverted (median frequency of 76.5% and 66.5%, respectively). IFNγ responses targeted the V2 loop for subjects who remained seronegative. HIV-1 gp120 envelope V2 loop-specific CD8 T-cell responses may help to protect against HIV-1 acquisition.

Background: Measles outbreaks are reported worldwide and pose a serious threat, especially to young unvaccinated infants. Early measles vaccination given to infants under 12 months of age can induce protective antibody levels, but the long-term antibody functionalities are unknown.Methods: Measles-specific antibody functionality was tested using a systems serology approach for children who received an early measles vaccination at 6-8 or 9-12 months, followed by a regular dose at 14 months of age, and children who only received the vaccination at 14 months. Antibody functionalities comprised complement deposition, cellular cytotoxicity, and neutrophil and cellular phagocytosis. We used Pearson's r correlations between all effector functions to investigate the coordination of the response.Results: Children receiving early measles vaccination at 6-8 or 9-12 months of age show polyfunctional antibody responses. Despite significant lower levels of antibodies in these early-vaccinated children, Fc effector functions were comparable with regular-timed vaccinees at 14 months. However, 3-year follow-up revealed significant decreased polyfunctionality in children who received a first vaccination at 6-8 months of age, but not in children who received the early vaccination at 9-12 months.Conclusions: Antibodies elicited in early-vaccinated children are equally polyfunctional to those elicited from children who received vaccination at 14 months. However, these antibody functionalities decay more rapidly than those induced later in life, which may lead to suboptimal, long-term protection. [ABSTRACT FROM AUTHOR]

SARS‐CoV‐2 specific T‐cell response has been associated with disease severity, immune memory and heterologous response to endemic coronaviruses. However, an integrative approach combining a comprehensive analysis of the quality of SARS‐CoV‐2 specific T‐cell response with antibody levels in these three scenarios is needed. In the present study, we found that, in acute infection, while mild disease was associated with high T‐cell polyfunctionality biased to IL‐2 production and inversely correlated with anti‐S IgG levels, combinations only including IFN‐γ with the absence of perforin production predominated in severe disease. Seven months after infection, both non‐hospitalised and previously hospitalised patients presented robust anti‐S IgG levels and SARS‐CoV‐2 specific T‐cell response. In addition, only previously hospitalised patients showed a T‐cell exhaustion profile. Finally, combinations including IL‐2 in response to S protein of endemic coronaviruses were the ones associated with SARS‐CoV‐2 S‐specific T‐cell response in pre‐COVID‐19 healthy donors' samples. These results could have implications for protective immunity against SARS‐CoV‐2 and recurrent COVID‐19 and may help for the design of new prototypes and boosting vaccine strategies. [ABSTRACT FROM AUTHOR]

Respiratory Syncytial Virus (RSV) is ubiquitous and re-infection with both subtypes (RSV/A and RSV/B) is common. The fusion (F) protein of RSV is antigenically conserved, induces neutralizing antibodies, and is a primary target of vaccine development. Insight into the breadth and durability of RSV-specific adaptive immune response, particularly to the F protein, may shed light on susceptibility to re-infection. We prospectively enrolled healthy adult subjects (n = 19) and collected serum and peripheral blood mononuclear cells (PBMCs) during the 2018–2019 RSV season. Previously, we described their RSV-specific antibody responses and identified three distinct antibody kinetic profiles associated with infection status: uninfected (n = 12), acutely infected (n = 4), and recently infected (n = 3). In this study, we measured the longevity of RSV-specific memory T cell responses to the F protein following natural RSV infection. We stimulated PBMCs with overlapping 15-mer peptide libraries spanning the F protein derived from either RSV/A or RSV/B and found that memory T cell responses mimic the antibody responses for all three groups. The uninfected group had stable, robust memory T cell responses and polyfunctionality. The acutely infected group had reduced polyfunctionality of memory T cell response at enrollment compared to the uninfected group, but these returned to comparable levels by end-of-season. The recently infected group, who were unable to maintain high levels of RSV-specific antibody following infection, similarly had decreased memory T cell responses and polyfunctionality during the RSV season. We observed subtype-specific differences in memory T cell responses and polyfunctionality, with RSV/A stimulating stronger memory T cell responses with higher polyfunctionality even though RSV/B was the dominant subtype in circulation. A subset of individuals demonstrated an overall deficiency in the generation of a durable RSV-specific adaptive immune response. Because memory T cell polyfunctionality may be associated with protection against re-infection, this latter group would likely be at greater risk of re-infection. Overall, these results expand our understanding of the longevity of the adaptive immune response to the RSV fusion protein and should be considered in future vaccine development efforts. [ABSTRACT FROM AUTHOR]

CD8+ T-cells play a crucial role in the control of HIV replication. HIV-specific CD8+ T-cell responses rapidly expand since the acute phase of the infection, and it has been observed that HIV controllers harbor CD8+ T-cells with potent anti-HIV capacity. The development of CD8+ T-cell-based vaccine against HIV-1 has focused on searching for immunodominant epitopes. However, the strong immune pressure of CD8+ T-cells causes the selection of viral variants with mutations in immunodominant epitopes. Since HIV-1 mutations are selected under the context of a specific HLA-I, the circulation of viral variants with these mutations is highly predictable based on the most prevalent HLA-I within a population. We previously demonstrated the adaptation of circulating strains of HIV-1 to the HLA-A*02 molecule by identifying mutations under positive selection located in GC9 and SL9 epitopes derived from the Gag protein. Also, we used an in silico prediction approach and evaluated whether the mutations found had a higher or lower affinity to the HLA-A*02. Although this strategy allowed predicting the interaction between mutated peptides and HLA-I, the functional response of CD8+ T-cells that these peptides induce is unknown. In the present work, peripheral blood mononuclear cells from 12 HIV-1+ HLA-A*02:01+ individuals were stimulated with the mutated and wild-type peptides derived from the GC9 and SL9 epitopes. The functional profile of CD8+ T-cells was evaluated using flow cytometry, and the frequency of subpopulations was determined according to their number of functions and the polyfunctionality index. The results suggest that the quality of the response (polyfunctionality) could be associated with the binding affinity of the peptide to the HLA molecule, and the functional profile of specific CD8+ T-cells to mutated epitopes in individuals under cART is maintained. [ABSTRACT FROM AUTHOR]

The research is performed within the linguistic stylistic approach which allows to determine the pragmatic foundation as existence of the ironic mode. The article is devoted to consideration of travelogue as a special type of text and to the evidence that irony is one of the universal features of travelogue as a text, which helps to define the borders between "own" and "other" and is aimed at perception of these borders. The material for analysis includes the texts of the 19 th and 20 th centuries, which vividly express the author's ironic attitude towards the reality. The functions of ironic mode in the Russian travelogue are revealed. The dominating role of the educating function is demonstrated. It is determined that ironic mode is characterized by polyfunctionality – simultaneous actualization of several functions, including: motivational (the motivation of readers to follow the author's travel as well as accept his ideas), expressive and contact-setting (as combination of contact initiation and harmonization of communication), aesthetic (making an object of description artistic and sensitive) functions. The research might continue with studying the ironic mode in travelogue of different times in a comparative aspect as well as analyzing the ironic mode within the peculiarities of the author's individual style.

Respiratory Syncytial Virus (RSV) is ubiquitous and re-infection with both subtypes (RSV/A and RSV/B) is common. The fusion (F) protein of RSV is antigenically conserved, induces neutralizing antibodies, and is a primary target of vaccine development. Insight into the breadth and durability of RSV-specific adaptive immune response, particularly to the F protein, may shed light on susceptibility to re-infection. We prospectively enrolled healthy adult subjects (n = 19) and collected serum and peripheral blood mononuclear cells (PBMCs) during the 2018–2019 RSV season. Previously, we described their RSV-specific antibody responses and identified three distinct antibody kinetic profiles associated with infection status: uninfected (n = 12), acutely infected (n = 4), and recently infected (n = 3). In this study, we measured the longevity of RSV-specific memory T cell responses to the F protein following natural RSV infection. We stimulated PBMCs with overlapping 15-mer peptide libraries spanning the F protein derived from either RSV/A or RSV/B and found that memory T cell responses mimic the antibody responses for all three groups. The uninfected group had stable, robust memory T cell responses and polyfunctionality. The acutely infected group had reduced polyfunctionality of memory T cell response at enrollment compared to the uninfected group, but these returned to comparable levels by end-of-season. The recently infected group, who were unable to maintain high levels of RSV-specific antibody following infection, similarly had decreased memory T cell responses and polyfunctionality during the RSV season. We observed subtype-specific differences in memory T cell responses and polyfunctionality, with RSV/A stimulating stronger memory T cell responses with higher polyfunctionality even though RSV/B was the dominant subtype in circulation. A subset of individuals demonstrated an overall deficiency in the generation of a durable RSV-specific adaptive immune response. Because memory T cell polyfunctionality may be associated with protection against re-infection, this latter group would likely be at greater risk of re-infection. Overall, these results expand our understanding of the longevity of the adaptive immune response to the RSV fusion protein and should be considered in future vaccine development efforts.

CD8+ T-cells play a crucial role in the control of HIV replication. HIV-specific CD8+ T-cell responses rapidly expand since the acute phase of the infection, and it has been observed that HIV controllers harbor CD8+ T-cells with potent anti-HIV capacity. The development of CD8+ T-cell-based vaccine against HIV-1 has focused on searching for immunodominant epitopes. However, the strong immune pressure of CD8+ T-cells causes the selection of viral variants with mutations in immunodominant epitopes. Since HIV-1 mutations are selected under the context of a specific HLA-I, the circulation of viral variants with these mutations is highly predictable based on the most prevalent HLA-I within a population. We previously demonstrated the adaptation of circulating strains of HIV-1 to the HLA-A*02 molecule by identifying mutations under positive selection located in GC9 and SL9 epitopes derived from the Gag protein. Also, we used an in silico prediction approach and evaluated whether the mutations found had a higher or lower affinity to the HLA-A*02. Although this strategy allowed predicting the interaction between mutated peptides and HLA-I, the functional response of CD8+ T-cells that these peptides induce is unknown. In the present work, peripheral blood mononuclear cells from 12 HIV-1+ HLA-A*02:01+ individuals were stimulated with the mutated and wild-type peptides derived from the GC9 and SL9 epitopes. The functional profile of CD8+ T-cells was evaluated using flow cytometry, and the frequency of subpopulations was determined according to their number of functions and the polyfunctionality index. The results suggest that the quality of the response (polyfunctionality) could be associated with the binding affinity of the peptide to the HLA molecule, and the functional profile of specific CD8+ T-cells to mutated epitopes in individuals under cART is maintained.

While corpus studies have shown that discourse connectives that convey the same coherence relation can display subtle differences, research on online discourse processing has only focused on a rather limited set of connectives. Yet, different connectives – for example, rare or polyfunctional ones – might elicit different reading patterns. In order to explore this assumption, we test the robustness of discourse processing for French native speakers by measuring the way they process causal and concessive sentences that are conveyed by either an appropriate or inappropriate connective. Throughout three experiments, we change important characteristics of the connectives: we first test frequently used connectives (Experiment 1), secondly less frequent ones (Experiment 2), and finally less frequent connectives that are polyfunctional and for which different functions clearly compete (Experiment 3). Our results show that the processing for incoherent items was affected for all connectives, however readers showed altered reading fluency when infrequent connectives were used. We conclude that discourse processing is quite robust and that readers are able to insert meaning conveyed by rare connectives while still showing the highest reading ease with frequent connectives.

While corpus studies have shown that discourse connectives that convey the same coherence relation can display subtle differences, research on online discourse processing has only focused on a rather limited set of connectives. Yet, different connectives – for example, rare or polyfunctional ones – might elicit different reading patterns. In order to explore this assumption, we test the robustness of discourse processing for French native speakers by measuring the way they process causal and concessive sentences that are conveyed by either an appropriate or inappropriate connective. Throughout three experiments, we change important characteristics of the connectives: we first test frequently used connectives (Experiment 1), secondly less frequent ones (Experiment 2), and finally less frequent connectives that are polyfunctional and for which different functions clearly compete (Experiment 3). Our results show that the processing for incoherent items was affected for all connectives, however readers showed altered reading fluency when infrequent connectives were used. We conclude that discourse processing is quite robust and that readers are able to insert meaning conveyed by rare connectives while still showing the highest reading ease with frequent connectives. [ABSTRACT FROM AUTHOR]

Copyright of Linguistische Treffen in Wrocław is the property of Oficyna Wydawnicza ATUT and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Background: Characterizing the longevity and quality of cellular immune responses to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) enhances understanding of coronavirus disease 2019 (COVID-19) immunity that influences clinical outcomes. Prior studies suggest SARS-CoV-2-specific T cells are present in peripheral blood 10 months after infection. Analysis of the function, durability, and diversity of cellular response long after natural infection, over a range of ages and disease phenotypes, is needed to identify preventative and therapeutic interventions.Methods: We identified participants in our multisite longitudinal, prospective cohort study 12 months after SARS-CoV-2 infection representing a range of disease severity. We investigated function, phenotypes, and frequency of T cells specific for SARS-CoV-2 using intracellular cytokine staining and spectral flow cytometry, and compared magnitude of SARS-CoV-2-specific antibodies.Results: SARS-CoV-2-specific antibodies and T cells were detected 12 months postinfection. Severe acute illness was associated with higher frequencies of SARS-CoV-2-specific CD4 T cells and antibodies at 12 months. In contrast, polyfunctional and cytotoxic T cells responsive to SARS-CoV-2 were identified in participants over a wide spectrum of disease severity.Conclusions: SARS-CoV-2 infection induces polyfunctional memory T cells detectable at 12 months postinfection, with higher frequency noted in those who experienced severe disease. [ABSTRACT FROM AUTHOR]

CD8 T cell memory offers critical antiviral protection, even in the absence of neutralizing antibodies. The paradigm is that CD8 T cell memory within the lung tissue consists of a mix of circulating TEM cells and non-circulating TRM cells. However, based on our analysis, the heterogeneity within the tissue is much higher, identifying TCM, TEM, TRM, and a multitude of populations which do not perfectly fit these classifications. Further interrogation of the populations shows that TRM cells that express CD49a, both with and without CD103, have increased and diverse effector potential compared with CD49a negative populations. These populations function as a one-man band, displaying antiviral activity, chemokine production, release of GM-CSF, and the ability to kill specific targets in vitro with delayed kinetics compared with effector CD8 T cells. Together, this study establishes that CD49a defines multiple polyfunctional CD8 memory subsets after clearance of influenza infection, which act to eliminate virus in the absence of direct killing, recruit and mature innate immune cells, and destroy infected cells if the virus persists.