Marta Massanella

IrsiCaixa Institute for AIDS Research | IRSICAIXA · Translational Research on Immunology and Aging

Background At least 5–10% of subjects surviving COVID-19 develop the post-COVID-19 condition (PCC) or “Long COVID”. The clinical presentation of PCC is heterogeneous, its pathogenesis is being deciphered, and objective, validated biomarkers are lacking. It is unknown if PCC is a single entity or a heterogeneous syndrome with overlapping pathophysio...

Most individuals acutely infected with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) exhibit mild symptoms. However, 10 to 20% of those infected develop long-term symptoms, referred to as post-coronavirus disease 2019 (COVID-19) condition (PCC). One hypothesis is that PCC might be exacerbated by viral persistence in tissue sanctuarie...

The elicitation of cross-variant neutralizing antibodies against SARS-CoV-2 represents a major goal for current COVID-19 vaccine strategies. Additionally, natural infection may also contribute to broaden neutralizing responses. To assess the contribution of vaccines and natural infection, we cross-sectionally analyzed plasma neutralization titers o...

The phenotype of the rare HIV-infected cells persisting during antiretroviral therapies (ART) remains elusive. We developed a single-cell approach that combines the phenotypic analysis of HIV-infected cells with near full-length sequencing of their associated proviruses to characterize the viral reservoir in 6 male individuals on suppressive ART. W...

Patients with solid tumors have been a risk group since the beginning of the SARS‐CoV‐2 pandemic due to more significant complications, hospitalizations, or deaths. The immunosuppressive state of cancer treatments or the tumor itself could influence the development of post‐vaccination antibodies. This study prospectively analyzed 89 patients under...

SARS-CoV-2 vaccination is the most effective strategy to protect individuals with haematologic malignancies against severe COVID-19, while eliciting limited vaccine responses. We characterized the humoral responses following 3 mo after mRNA-based vaccines in individuals at different plasma-cell disease stages: monoclonal gammopathy of undetermined...

HIV latent infection may be associated with disrupted viral RNA sensing, interferon (IFN) signaling, and/or IFN stimulating genes (ISG) activation. Here, we evaluated the use of compounds selectively targeting at the inhibitor of nuclear factor-κB (IκB) kinase (IKK) complex subunits and related kinases (TBK1) as a novel pathway to reverse HIV-1 lat...

Background: Prophylactic vaccination has proven to be the most effective strategy to fight the COVID-19 pandemic. Methods: This was a prospective observational cohort study involving 30 predominantly antibody deficiency disorders (ADD)-afflicted adult patients on immunoglobulin replacement therapy vaccinated with three doses of the mRNA-1273 COV...

Mass vaccination campaigns reduced COVID-19 incidence and severity. Here, we evaluated the immune responses developed in SARS-CoV-2-uninfected patients with predominantly antibody-deficiencies (PAD) after three mRNA-1273 vaccine doses. PAD patients were classified based on their immunodeficiency: unclassified primary antibody-deficiency (unPAD, n=9...

We describe a case of a patient who experienced recurrent COVID-19 pneumonia over a period of 123 days. Neither remdesivir nor convalescent plasma were temporally associated with viral clearance or increased plasma neutralization capacity. Antibody levels remained low until day 151 and cellular immunity increased overtime with no viral evolution.

The phenotype of the rare HIV-infected cells persisting during ART remains elusive. We developed a single-cell approach that combines the phenotypic analysis of HIV-infected cells with near full-length sequencing of their associated proviruses. Individual cells carrying clonally expanded identical proviruses displayed very diverse phenotypes, indic...

The persistence of latent HIV reservoirs allows for viral rebound upon antiretroviral therapy interruption, hindering effective HIV-1 cure. Emerging evidence suggests that modulation of innate immune stimulation could impact viral latency and contribute to the clearing of HIV reservoir. Here, the latency reactivation capacity of a subclass of selec...

Limited data exists on SARS-CoV-2 sustained-response to vaccine in patients with rheumatic diseases. This study aims to evaluate neutralizing antibodies (nAB) induced by SARS-CoV-2 vaccine after 3 to 6 months from administration in Systemic Lupus Erythematosus (SLE) patients, as a surrogate of sustained-immunological response. This cross-sectional...

The diagnosis of the post-COVID condition is usually achieved by excluding other diseases; however, cognitive changes are often found in the post-COVID disorder. Therefore, monitoring and treating the recovery from the post-COVID condition is necessary to establish biomarkers to guide the diagnosis of symptoms, including cognitive impairment. Our s...

The role of T cells in the control of SARS-CoV-2 infection has been underestimated in favor of neutralizing antibodies. However, cellular immunity is essential for long-term viral control and protection from disease severity. To understand T-cell immunity in the absence of antibody generation we focused on a group of SARS-CoV-2 Non-Seroconvertors (...

Background: SARS-CoV-2 vaccination is the most effective strategy to protect older residents of long-term care facilities (LTCF) against severe COVID-19, but primary vaccine responses are less effective in older adults. Here, we characterised the humoral responses of institutionalised seniors 3 months after they had received the mRNA/BNT162b2 vacc...

Purpose Mass vaccination campaigns have reduced the incidence and severity of COVID-19. However, there is limited information about how patients with predominantly antibody-deficiencies (PAD) respond to COVID-19 vaccination. Here, we evaluated humoral and cellular responses developed in SARS-CoV-2-naïve PAD individuals after three mRNA-1273 vaccine...

Seroconversion panels were collected before and after vaccination with three COVID-19 vaccines: two mRNA vaccines (mRNA-1273 and BNT-162b2) and one adenovirus vector vaccine (Ad26.COV2.S). The panels were tested for antibody activity by chemiluminescent immunoassay, ELISA and one was tested in a pseudovirus neutralization assay. Participants positi...

Current diagnostics of severe acute respiratory syndrome coronavirus 2 (SARS-CoV2) infection heavily rely on reverse transcription-polymerase chain reaction (RT-PCR) or on rapid antigen detection tests. The former suffers from long time-to-result and high cost while the latter from poor sensitivity. Therefore, it is crucial to develop rapid, sensit...

To understand the determinants of long-term immune responses to SARS-CoV-2 and the concurrent impact of vaccination and emerging variants, we follow a prospective cohort of 332 COVID-19 patients over more than a year after symptom onset. We evaluate plasma neutralizing activity using HIV-based pseudoviruses, expressing the spike of different SARS-C...

SARS-CoV-2 vaccination is the most effective strategy to protect patients with haematologic malignancies against severe COVID-19, but primary vaccine responses are less effective in this population. Here, we characterized the humoral responses following 3 months after mRNA-based vaccines in patients at different stages of the same plasma cell disea...

COVID-19 pathophysiology is currently not fully understood, reliable prognostic factors remain elusive, and few specific therapeutic strategies have been proposed. In this scenario, availability of biomarkers is a priority. MS-based Proteomics techniques were used to profile the proteome of 81 plasma samples extracted in four consecutive days from...

Inappropriate sinus tachycardia (IST) is a common observation in patients with post-COVID-19 syndrome (PCS) but has not yet been fully described to date. To investigate the prevalence and the mechanisms underlying IST in a prospective population of PCS patients. Consecutive patients admitted to the PCS Unit between June and December 2020 with a res...

Background: SARS-CoV-2 vaccination is the most effective strategy to protect elders living in long-term care facilities (LTCF) against severe COVID-19, but primary vaccine responses are less effective in older adults. Here, we characterized the humoral responses following 3 months after mRNA/BNT162b2 vaccine in institutionalized elders. Methods: Pl...

Angiotensin converting enzyme 2 (ACE2) is a host ectopeptidase and the receptor for the SARS-CoV-2 virus, albeit virus-ACE2 interaction goes far beyond viral entry into target cells. Controversial data exists linking viral infection to changes in ACE2 expression and function, which might influence the subsequent induction of an inflammatory respons...

Background: Understanding the determinants of long-term immune responses to SARS-CoV-2 and the concurrent impact of vaccination and emerging variants of concern will guide optimal strategies to achieve global protection against the COVID-19 pandemic. Methods: A prospective cohort of 332 COVID-19 patients was followed beyond one year. Plasma neutral...

Background SARS-CoV-2 reinfections have been reported; however, most cases are milder than the primary infection. We report the first case of a life-threatening critical presentation of a SARS-CoV-2 reinfection. Case presentation A 62-year male from Palamós (Spain) suffered a first mild COVID-19 episode in March 2020, confirmed by 2 independent SA...

With the spread of new variants of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), there is a need to assess the protection conferred by both previous infections and current vaccination. Here we tested the neutralizing activity of infected and/or vaccinated individuals against pseudoviruses expressing the spike of the original SARS-Co...

Funding Acknowledgements Type of funding sources: None. Background Persistent symptoms after the acute phase of SARS-CoV-2 infection are referred to as "post-COVID-19 syndrome" (PCS), with a reported incidence ranging between 35% and 87%. Fatigue, palpitations and exercise intolerance are common complains among PCS patients in whom unexplained sin...

The role of T cells in the control of SARS-CoV-2 infection has been underestimated in favor of neutralizing antibodies. However, cellular immunity is essential for long-term viral control and protection from disease severity. To understand T-cell immunity in the absence of antibody generation we focused on a group of SARS-CoV-2 Non-Seroconvertors (...

To assess the potential impact of predominant circulating SARS-CoV-2 variants on neutralizing activity of infected and/or vaccinated individuals, we analyzed neutralization of pseudoviruses expressing the spike of the original Wuhan strain, the D614G and B.1.1.7 variants. Our data show that parameters of natural infection (time from infection and i...

Background Chronic inflammation and residual HIV transcription persist in people living with HIV (PLWH) receiving antiretroviral therapy (ART), thus increasing the risk of developing non-AIDS co-morbidities. The mechanistic target of rapamycin (mTOR) is a key regulator of cellular metabolism and HIV transcription, and therefore represents an intere...

Background Early antiretroviral therapy (ART) initiation (ie, within 3 months of infection) limits establishment of the HIV reservoir. However, the effect of early ART initiation on the long-term dynamics of the pool of infected cells remains unclear. Methods In this longitudinal analysis, we included cisgender men who have sex with men (MSM) and...

Background Understanding mid-term kinetics of immunity to SARS-CoV-2 is the cornerstone for public health control of the pandemic and vaccine development. However, current evidence is rather based on limited measurements, losing sight of the temporal pattern of these changes. Methods We conducted a longitudinal analysis on a prospective cohort of...

The protective effect of neutralizing antibodies in SARS-CoV-2 infected individuals is not yet well defined. To address this issue, we have analyzed the kinetics of neutralizing antibody responses and their association with disease severity. Between March and May 2020, the prospective KING study enrolled 72 COVID-19+ participants grouped according...

Relative control of HIV-1 infection has been linked to genetic and immune host factors. In this study, we analyzed 96 plasma proteome arrays from chronic untreated HIV-1-infected individuals using the classificatory random forest approach to discriminate between uncontrolled disease (plasma viral load [pVL] >50,000 RNA copies/ml; CD4 counts 283 cel...

Understanding mid-term kinetics of immunity to SARS-CoV-2 is the cornerstone for public health control of the pandemic and vaccine development. However, current evidence is rather based on limited measurements, thus losing sight of the temporal pattern of these changes 1–6 . In this longitudinal analysis, conducted on a prospective cohort of COVID-...

There is an urgent need to elucidate the molecular mechanisms underlying the transmissibility and pathogenesis of SARS-CoV-2. ACE2 is a host ectopeptidase with well-described anti-inflammatory and tissue protective functions and the receptor for the virus. Understanding SARS-CoV-2-ACE2 interaction and the expression of antiviral host genes in early...

Background: Early antiretroviral therapy (ART) restricts the size of the HIV reservoir in infants. However, whether antiretroviral (ARV) prophylaxis given to exposed vertically infected children exerts similar effects remains unknown. Methods: We measured total and integrated HIV DNA, as well as the frequency of CD4 T-cells producing multiply-sp...

The persistence of replication-competent HIV reservoirs in people living with HIV (PLWH) receiving antiretroviral therapy (ART) is a barrier to cure. Therefore, their accurate quantification is essential for evaluating the efficacy of new therapeutic interventions and orienting the decision to interrupt ART. Quantitative viral outgrowth assays (QVO...

Latently infected CD4 lymphocytes preclude cure of HIV infection, even with the most effective antiretroviral therapy. The replication competent latent HIV reservoir has been quantified with the terminal dilution quantitative viral outgrowth assay, which induces virus propagation in CD4⁺ T cell culture supernatants following cellular activation. Ef...

The role of circulating monocytes as persistent HIV reservoirs during ART is still controversial. Several studies have reported persistent infection of monocytes in virally suppressed individuals; however, others failed to detect HIV in this subset. These discrepancies are likely explained by the diversity of the methods used to isolate monocytes a...

Chronic viral infections such as with HIV and CMV last a lifetime and can continually antagonize the immune system. Both viruses are associated with higher expression of inflammation markers, and recent evidence suggests that CMV may complicate efforts to deplete HIV reservoirs. Our group and others have shown that CMV shedding is associated with a...

Quantitative viral outgrowth assays (QVOA) use limiting dilutions of CD4⁺ T cells to measure the size of the latent HIV-1 reservoir, a major obstacle to curing HIV-1. Efforts to reduce the reservoir require assays that can reliably quantify its size in blood and tissues. Although QVOA is regarded as a “gold standard” for reservoir measurement, litt...

Laboratory protocols. (PDF)

QVOA well configuration. (XLSX)

Experimental setup, showing lab and batch assignment of each aliquot. (XLSX)

Complete experimental dataset. (XLSX)

Performance of MCMC estimation using the model with positive aliquot- and batch-level variation and HC(0,2) prior, in single-lab simulation. (PDF)

Performance of MCMC estimation using the ensemble model and HC(0,4) prior, in multi-lab simulation, as in Table 2. (PDF)

Performance of MCMC estimation assuming a true random effect parameter of zero, using HC(0,1) prior, in multi-lab simulation. (PDF)

Summary of model fits. Parameter distributions of the ensemble model are reported as weighted quantiles of 8000 posterior samples, and the number of samples in the discrete ensembles are shown (see Methods: “Markov-chain Monte Carlo estimation”). Analysis is shown for eight different subsets of the data: three subsets including multiple labs, and f...

Pairwise comparisons of assay accuracy for higher IUPMs, assuming that each assay has equal claim to biological truth (σc applied to both assays). Method and format match S12 Table. (PDF)

Performance of MCMC estimation using the ensemble model and HC(0,2) prior, in multi-lab simulation, as in Table 2. (PDF)

Performance of MCMC estimation using the ensemble model and HC(0,3) prior, in multi-lab simulation, as in Table 2. (PDF)

Pairwise comparisons of assay accuracy for lower IUPMs, assuming that each assay has equal claim to biological truth (σc applied to both assays). Batch variation-free ensemble estimates of parameters were used in simulations. Each entry shows median improvement of row versus column assay (median and 95% CI sampling from ensemble parameter distribut...

Pairwise comparisons of assay accuracy for lower IUPMs, treating the JHU assay as gold standard (2×σc applied to other assays). Method and format match S12 Table. (PDF)

Pairwise comparisons of assay accuracy for higher IUPMs, treating the JHU assay as gold standard (2×σc applied to other assays). Method and format match S12 Table. (PDF)

Simulated results of latency reduction trials, maximum likelihood estimation to compare pre- and post-treatment data. (PDF)

Sensitivity analysis of of aliquot-level, batch-level, and lab-level variation. (TIF)

Scatterplot of estimates of excess variation at the aliquot, batch, and lab levels, expressed as the natural log of fold-change (parameters σa, σb, σc). Panels show robust linear regression and Wald test p-values (null model = zero slope) of 1000 samples from the ensemble posterior. (TIF)

The phenotypic characterization of the cells in which HIV persists during antiretroviral therapy (ART) remains technically challenging. We developed a simple flow cytometry-based assay to quantify and characterize infected cells producing HIV proteins during untreated and treated HIV infection. By combining two antibodies targeting the HIV capsid i...

MFI of p24 antibodies following stimulation. Comparison of the MFI of the two p24 antibodies (p24 28B7-APC and p24 KC57-PE) in the presence or absence of stimulation with PMA/ionomycin in samples from 6 untreated individuals. The MFI of p24 antibodies was measured within the p24+ gate (p24 KC57+/p24 28B7+). (TIF)

Comparison of two permeabilization buffers for the detection of p24-producing cells by HIV-Flow. Dot plots showing the detection of p24+ cells in 2 samples, using 2 experimental conditions. Purified CD4 T cells from a viremic individual were rested for 18 hours, while purified CD4 T cells from an ART-suppressed individual were stimulated with PMA/i...

Markers showing significant changes of expression following stimulation. (A) Representative dot plots showing the levels of expression of CXCR3/CCR4/CCR6 after 24h of resting or after 24h of stimulation with PMA/ionomycin + BFA in one representative ART-suppressed individual. (B) As in A) for CXCR5 and CD25. (C) As in A) for CD3 and CD4. Of note, t...