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Introduction: Epstein-Barr virus (EBV) infection precedes signs of multiple sclerosis (MS) pathology and cross-reactive antibodies might link EBV infection mechanistically to CNS autoimmunity. Objective(s): As an altered immune reaction against EBV antigens in T cells of MS patients has been suggested, we queried deep, peripheral blood T-cell receptor beta chain (TCRbeta) repertoires of 1395 MS patients, 887 controls, and 35 monozygotic, MS-discordant twin pairs for multimerconfirmed, viral antigen-specific TCRbeta sequences. Aim(s): Quantification of HLA-matched EBV-specific, CMV-specific, Influenza A virus-specific, and SARS-CoV-2-specific TCRbeta sequences in MS patients, controls, and COVID-19 patients. Result(s): We detected higher numbers of MHC-I restricted EBVspecific TCRbeta sequences in MS patients, and validated this with independent cohorts and sequencing methods. Genetic as well as early environmental factors could be excluded by validation in diseased siblings of monozygotic twin pairs discordant for MS. Therapeutic blockade of VLA-4-mediated T-cell extravasation amplified this observation, while interferon beta treatment and B-cell depletion did not modulate occurrence of EBV-specific T cells. EBV-specific CD8+ T cells were characterized as effectormemory cells in peripheral blood and cerebrospinal fluid of healthy controls. In MS patients, the cerebrospinal fluid also contained EBV-specific central-memory CD8+ T cells, suggesting recent priming. Conclusion(s): MS is not only preceded by EBV infection, but also associated with a broader EBV-specific TCR repertoire, which would be consistent with an ongoing anti-EBV immune reaction in MS patients.
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Introduction: Anti-SARS-CoV2 vaccination induces specific Tand B-cell responses in healthy subjects (HS). In MS patients treated with anti-CD20 drugs, the antibody response is reduced or absent, whereas specific T-cell responses are maintained. It is not known whether and how vaccination affects innate responses mediated by natural killer (NK) cells in HS and in MS patients treated with anti-CD20 drugs. Objective(s): To evaluate whether and how NK cells contribute to the immune response following anti-SARS-CoV2 vaccination in HS and in ocrelizumab-treated MS patients Aims: The aims of this work were: 1) to evaluate the effects of anti-SARS CoV2 vaccination on the phenotype of NK cells from HS and from ocrelizumab-treated MS patients and 2) to evaluate how peptides from the SARS-CoV2 spike protein affect NK cell responses before and after anti-SARS-CoV2 vaccination. Method(s): We enrolled 21 MS patients treated with ocrelizumab and 20 HS. Peripheral blood mononuclear cells (PBMCs) were isolated from peripheral blood and stored under liquid nitrogen. Thawed PBMCs were cultured overnight in presence/absence of SARS-CoV2 peptides or peptides from the cytomegalovirus (CMV), with/without activating cytokines. Phenotype of NK cells through a 13-marker flow cytometry panel and intracellular production of IFN-gamma were evaluated after culture. Result(s): Findings: 1) Vaccination increased the proportion of CD56dim NK cells in HS and MS patients. CD56posCD16neg NK cells, more abundant in MS patients before vaccination, decreased thereafter. Lower pre-vaccination activation capability of NK cells from MS patients compared to HS in response to stimulus with cytokines was reverted by vaccination. 2) Before vaccination, peptides from the SARS-CoV2 protein downregulated the production of IFN- gamma from NK cells of HS, but not ocrelizumabtreated MS patients, who had significantly lower baseline IFN-gamma NK cells 3) After vaccination, peptides from the SARS-CoV2 protein did not affect the production of IFN- gamma from NK cells of HS. Conclusion(s): The results of this work demonstrate anti-SARSCoV2 vaccination increases the proportion of effector CD56dim NK cells in HS and ocrelizumab-treated patients. Spike peptides inhibit the function of NK cells from HS before, but not after vaccination. Such phenomenon may contribute to the pathogenicity of SARS-CoV2 in unvaccinated subjects.
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Introduction: In patients with Multiple Sclerosis (pwMS), specific disease modifying treatments (DMTs) may compromise immune response following SARS-CoV-2 vaccination. Limited information is available, whether levels of anti-SARS-CoV-2 antibodies are linked to the risk of breakthrough infections in pwMS. Objective(s): To determine the rate of Omicron breakthrough infection and severity of COVID-19 in a cohort of MS patients treated with different DMTs and to estimate the impact of SARSCoV- 2-specific antibody level on breakthrough infection risk. Method(s): This study is nested within the Swiss MS Cohort, a nationwide multicenter study that has recruited 1585 pwMS. Patients who received two doses of SARS-CoV-2 vaccines before Omicron became the dominant variant in Switzerland on Dec-15, 2021 and had a follow-up thereafter were included. Data on SARS-CoV-2 infections, severity of COVID-19 according to the WHO scale and SARS-CoV-2 vaccines were collected by questionnaires. Anti-SARS-CoV-2-S antibody levels were measured after the second vaccine dose. Incidence of infections grouped by antibody level after second vaccination was visualized using Kaplan-Meier curves. Cox regression models were used to estimate the impact of antibody levels on the hazard of breakthrough infection during follow-up. Result(s): 242 pwMS (median age 49y [39,58], 162 (67%) female, 36 (15%) with progressive disease, median EDSS 2.5 [1.5,4.0]) were included. 22 (9%) had SARS-CoV-2 infection and 137 (57%) at least one additional vaccine dose prior to Omicron start. Since then, 57 breakthrough infections were reported. Severity of breakthrough disease on WHO scale ranged from 1-10: 7 were asymptomatic, 46 were symptomatic as outpatients, 3 were hospitalized and 1 died. Patients with antibody levels >150U/ml (n = 95, 39%) after second vaccination had a 64% reduced risk for Omicron breakthrough-infection compared to patients with antibody levels <0.7U/ml (n = 81, 33%) (HR 0.36, 95%CI=0.18- 0.71, p<0.01). This effect was maintained after adjustment for DMT at vaccination and time since second vaccination Conclusion(s): Humoral immune response after second SARSCoV- 2 vaccination is associated with Omicron breakthrough infection rate, a finding contrasting general populations, where antibody levels seem to have little impact on protecting from Omicron infection. Most breakthrough infections in our cohort were mild. Analyses on the effect of booster vaccinations on serology and infection rates will follow.
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Background: Attenuated antibody and robust T-cell responses following SARS-CoV-2 vaccines have been reported in people with multiple sclerosis (PwMS) treated with ocrelizumab (OCR). Factors influencing humoral and cellular responses and how these relate to clinical protection are not well understood. Aim(s): To investigate the effect of OCR on antibody and T-cell responses to SARS-CoV-2 vaccines, and characterise clinical outcomes of breakthrough COVID-19 infections in PwMS. Method(s): Patients participating in three ongoing Phase IIIb studies evaluating the effectiveness and safety of OCR, who decided to receive a SARS-CoV-2 vaccine as part of national immunisation programmes, were invited to undergo optional exploratory assessment. Antibody responses to SARS-CoV-2 spike and nucleocapsid proteins were assessed using the Elecsys electrochemiluminescence assay. Interferon-gamma ELISpot (ImmunoSpot) was used to detect SARS-CoV-2-specific T cells against 4 different peptide pools of the spike protein. Vaccine breakthrough cases were defined as suspected or laboratory-confirmed COVID-19 infections occurring >=14 days after completion of the recommended primary immunisation schedule. Result(s): Up to Sept 2021, 111 patients provided samples for assessment of response to SARS-CoV-2 vaccination. Mean (SD) age was 41.7 (11.8) years;63.1% were female;72.1% had relapsing- remitting MS, 15.3% secondary progressive MS and 12.6% primary progressive MS.Most patients had been on treatment with OCR for at least 2 years. Mean (SD) of 78.5 (41.4) days elapsed between the first vaccine dose and last OCR infusion. 94/111 patients received an mRNA vaccine and 15 patients an adenoviral vaccine. Most samples were collected within 80 days following completion of the primary vaccine schedule. Antibody response was detected in 22/103 (21%) patients and T-cell responses to >=1 of the 4 different peptide pools in 83/95 (87%) patients. Four breakthrough infections were reported. Conclusion(s): In preliminary analysis, frequency of patients with an antibody and T-cell response were in line with published data for PwMS on OCR. Expanded analyses with a larger set of patients (approximately 450 patients, up to March 2022), including longitudinal responses, booster data, differences between vaccine platforms and assessment of factors that may affect immune responses will be presented. Correlations between level of immune responses and breakthrough infection diagnosis and severity will be explored.
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Background: Patients on OCR have attenuated antibody, but largely intact T-cell responses to COVID-19 vaccination. Little is known about durability of post-vaccine responses in OCR-treated patients. Objective(s): To examine antibody and cellular responses to mRNA COVID-19 vaccines (Pfizer, BioNTech/Moderna) in Ocrelizumab (OCR)-treated MS patients over 24-week period. Method(s): MS patients on OCR were recruited from NYU (New York City) and Rocky Mountain at CU (Denver) MS Centers. Antibody responses to SARS-CoV-2 spike proteins were assessed with multiplex bead-based (MBI) immunoassays, and cellular responses to SARS-CoV-2 Spike protein with ELISpot and activation induced marker (AIM) panel in a Cytek Aurora full-spectrum flow cytometry platform. Data on samples collected pre-vaccine and 4-, 12-, 24-weeks post 2-doses and 4-, 12-weeks post-third dose will be presented. Result(s): 40/61 enrollees (age 38.3+/-10.9;77.5% female;57.5% non-white) had 24-week post-vaccination data and 9 patients had 4-week post 3rd dose data. Antibody response increased from prevaccine level of 972.0 U/mL to 6307.4 U/mL at week-4 (p=0.0002), then decreased to 4633.8 u/mL at week-12 (26% decrease from week-4, p=0.1377), and further to 2878.4 u/mL at week-24 (37% decrease from week-12, p value=0.109). Spikespecific IFNgamma T-cell responses by ELIspot were 125.7 SFU/106 cells pre-vaccine, increased to 362.9 SFU/106 cells at week-4 (p=0.009), then to 511.5 SFU/106 cells at week-12 (40.9% increase relative to 4-week time-point, p=0.8474), and remained elevated at 501.7 SFU/106 cells at week-24 (p=0.7393, 1.9% compared to week 12). 4-week post 3rd dose, Ab level increased to 5094.8 U/mL (189.9% compared to pre-3rd dose, p =0.076) and IFNgamma responses to 1253.3 SFU/106 cells (484.5% increase, p=0.037). Conclusion(s): Antibody responses to 2-series vaccine peaked at 4 weeks and trended downward thereafter, while cellular responses were sustained at 24 weeks. Third-dose resulted in marked increases in both antibody and T-cell responses 4-weeks. Expanded analyses, including in-depth immunophenotyping and 12-week post 3rd vaccination responses will be presented.
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Objective: To compare humoral and cellular responses to COVID-19 vaccines in 400 consecutive MS patients who were on Ocrelizumab ('OCR') and other disease-modifying therapies ('nonOCR') at the time of vaccination. Background: Peripheral B-cell depletion with anti-CD20 therapies, attenuates humoral responses to vaccines, but less is known about cellular responses. Design/Methods: Consecutive MS patients from NYU MS Care Center were invited to participate if they completed COVID-19 vaccination ≥6 weeks previously. Immune testing included anti-spike RBD antibody (Elecsys Anti-SARS-CoV-2) (Roche Diagnostics);multiepitope bead-based immunoassays (MBI) of antibody-responses to SARS-COV-2 spike proteins (threshold of 'positivity'was chosen as 2 SD below non-OCR mean);T-cell responses to SARSCoV-2 Spike protein using IFNγ enzyme-linked immune-absorbent spot (Invitrogen) and TruCulture (Myriad RBM) assays;high dimensional immunophenotyping;live virus immunofluorescence-based microneutralization assay. Results: Antibody and T cell data was available on 145/355 patients enrolled to date (mean age: 40.0 years;75% female;48% non-white;39% on OCR;12% with prior COVID-19 infection;vaccines: 58% Pfizer/BioNTech, 36% Moderna and 6% Johnson&Johnson;median vaccine-tosample time: 93 (+/-32) days). In OCR, Elecsys Anti-SARS-CoV-2 Ab titers were detected in 30/63 (48%;mean antibody titer in log scale: 1.63) and in non-OCR - in 78/81 (96%, mean Ab titer in log scale: 2.83;p<0.0001). In OCR, antibody response by MBI were detected in 41/57 (72%, mean level in log scale: 3.09) and in non OCR - in 68/72 (94%, mean level in log scale: 4.08;p<0.001). Neutralizing antibodies were detected in 10/42 (38%) of OCR and 24/43 (56%) of non-OCR (p=0.1). T-cell activation based on induced IFNg secretion (TruCulture) was observed in 50/64 (78%) OCR and 43/81 (53%) non-OCR (p=0.002). Conclusions: Preliminary results suggest robust vaccine-specific T-cell immune response to SARS-CoV2 vaccines in B-cell depleted patients, but markedly attenuated antibody responses. Final results of pre-planned multivariable analyses stratified by DMT class and high-dimensional immunophenotyping will be presented.
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Objective: To examine antibody and T-cell responses to mRNAplatform COVID-19 vaccines in Ocrelizumab-treated MS patients over a 12-month period. Introduction: B-cell depletion with Ocrelizumab attenuates humoral responses to vaccines. The kinetics of humoral and cellular immune responses to COVID-19 vaccines in B-cell depleted MS patients has not been reported. Methods: VIOLA (NCT04843774) is an open-label, observational study enrolling 60 MS patients on Ocrelizumab from NYU and Rocky Mountain at the University of Colorado MS Centers. First vaccine dose occurred ≥2 weeks after ocrelizumab infusion;second-dose ≥8 weeks before the next infusion. Antibody responses to SARS-COV-2 spike proteins were assessed with Elecsys Anti-SARS-CoV-2 (Roche Diagnostics) and multiplex bead-based immunoassays. T-cell responses to SARS-CoV-2 Spike protein were assessed with IFNγ ELISpot (Invitrogen) and TruCulture (Myriad RBM) and high-dimensional immunophenotyping. Samples are collected pre-vaccination and at 4, 12, 24, and 48-weeks post-vaccination. Results: As of 7/15/2021, 52 subjects have been enrolled (39.7±10.0 years;73% female;47% non-white), of whom 47 were fully vaccinated (85% Pfizer, 15% Moderna). Anti-spike RBD antibody (Elecsys Anti-SARS-CoV-2) were available for pre- and post-vaccine timepoints for 15 patients. Pre-vaccine, 1/15 (7%) patients had detectable titers, while at 4-weeks postvaccine, 10/15 (66%) patients had detectible titers (mean for positives: 1189 U/ml;5 patients had positive titers <25 U/ml). T-cell activation based on induced IFNγ secretion (TruCulture) at baseline and 4-week post-vaccine timepoints were available for 13 patients, of whom 12 (92%) were increased (mean pre-vaccine: 24 pg/ml;mean post-vaccine: 366 pg/ml, two-tailed t-test, p=0.0032). Conclusions: This prospective study of humoral and cellular immune responses to COVID-19 vaccines in Ocrelizumab-treated patients will generate data to help guide management of MS patients on anti-CD20 therapies. Early results suggest that 4-weeks post-vaccination nearly all Ocrelizumab-treated MS patients develop T-cell immunity and two-thirds showed evidence of humoral response. Additional 4-week and 12-week post-vaccination data will be presented.
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Objective: To compare humoral and T-cell responses to COVID- 19 vaccines in 400 MS patients who were on Ocrelizumab ('OCR') v. other disease-modifying therapies ('non-OCR') at the time of vaccination. Introduction: Peripheral B-cell depletion with anti-CD20 therapies attenuates humoral responses to vaccines. Whether immune responses to COVID-19 vaccines differ between B-cell depleted and non-B cell depleted MS patients is not known. Methods: Consecutive MS patients from NYU MS Care Center were invited to participate if they completed COVID-19 vaccination ≥6 weeks previously. Immune testing included anti-spike RBD antibody (Elecsys Anti-SARS-CoV-2) (Roche Diagnostics);multiplex bead-based immunoassays of antibody-responses to SARS-COV-2 spike proteins;T-cell responses to SARS-CoV-2 Spike protein using IFNγ enzyme-linked immune-absorbent spot (Invitrogen) and TruCulture (Myriad RBM) assays;high dimensional immunophenotyping;and live virus immunofluorescencebased microneutralization assay. Results: As of 7/15/2021, 105 MS subjects were enrolled (mean age: 40.5 years;76% female;41% non-white;38% on OCR;12% with prior COVID-19 infection). 95% were fully vaccinated with mRNA vaccines (Pfizer/Moderna);5% - with adenovirus-based vaccine (Johnson&Johnson). Median time from sample collection to last vaccine was 79 days. Positive Elecsys Anti-SARS-CoV-2 Ab titers post-vaccine were detected in 11/37 (30%) in OCR (mean level: 702 U/mL among seropositives) and 54/54 (100%) patients in non-OCR (mean level: 2310 U/mL;p<0.0001). Positive response by multiplex assay (threshold of 'positive' defined as 2 SD below the mean for the non-OCR) were detected in 10/27 (37%) OCR and 29/31 (94%) non-OCR (p<0.00001). T-cell activation based on induced IFNγ secretion (TruCulture) was detected in 20/25 (80%) OCR and 16/19 (84%) non-OCR patients (p=0.71). Conclusions: Preliminary results suggest robust T-cell immune response to SARS-CoV2 vaccines in approximately 80% of both OCR and non-OCR MS patients. Antibody responses were markedly attenuated in OCR compared to non-OCR group. Updated results will be presented.
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Introduction: The SARS-CoV-2 pandemic has raised, among others, a particular concern for people taking immune-suppressants. The Italian MS Foundation (FISM), Neuroimmunology Association (AINI), Neurological Society (SIN), and MS Registry have constituted an Alliance to tackle these issues. In the field of multiple sclerosis, several reports have suggested a higher risk of infection and an increased severity of the disease in persons treated with anti-CD20 monoclonal antibody. Serological investigations, showing a blunted production of anti-SARS-CoV-2 antibodies, questioned the usefulness of vaccination in these subjects, without, however, considering T cell responses. Objectives and Aims: To investigate antiviral T cell responses after infection with SARS-CoV-2 in persons with MS (pwMS) treated with Ocrevus.Control groups include pwMS treated with Ocrevus without SARS-CoV-2 infection, persons without MS with SARS-CoV-2 infection, and healthy individuals vaccinated or not with BNT16b2. Methods: Blood samples were collected and processed to isolate PBMCs, that were then stored frozen. PBMCs were stimulated with SARS-CoV-2 peptide pools and T cell reactivity was assessed by ELISPOT for IFNg detection, and by multiparametric FACS analyses for assessment and characterization of T cell activation. Results: ELISPOT assay against the spike and the N protein of SARS-CoV-2 displayed specific T cell reactivity in 80% pwMS treated with Ocrevus and infected by SARS-CoV-2, similar to infected persons without MS. FACS analysis following stimulation with SARS-CoV-2 peptide pools, showed the presence of activation-induced markers (AIM) in both CD4 and CD8 T cell subsets in 96% and 92% of these individuals, respectively. CD4 AIM+ cells were mostly central and effector memory cells, while CD8 cells were largely CD45RA+ terminally differentiated effectors (TEMRA) and poised for cytotoxicity, with a significant fraction of naïve cells. Within naïve AIM+ CD4 and CD8 cells we detected memory stem cells, suggesting the acquisition of long-term memory and protection from reinfection. COVID-19- recovered pwMS treated with Ocrevus had T cell responses comparable to healthy individuals vaccinated with BNT162b2, particularly concerning the ability to produce cytokines. Conclusions: B-cell depletion using Ocrevus does not impair the development of anti-SARS-CoV-2 T cell responses. Multistakeholder initiatives are mandatory to rapidly obtain unbiased clinically crucial information.
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Background: COVID-19 disease course in MS has been described in various cohorts. Limited data is available on humoral immune responses following SARS-CoV-2 infection and vaccination. Objectives: To determine the rate of confirmed SARS-CoV-2 infection and severity of COVID-19 in a cohort of MS patients and to quantify SARS-CoV-2-specific antibody response. Methods: The study is nested within the Swiss MS Cohort, a nationwide multicenter study that has recruited 1504 persons with MS (pwMS) since 2012. PCR-confirmed SARS-CoV-2 infections, severity of COVID-19 according to the WHO clinical progression scale and immunizations with SARS-CoV-2 vaccines were captured by questionnaires used for interviews every 6 or 12 months. Anti-SARS-CoV-2 spike protein and nucleocapsid antibody levels will be determined by electrochemiluminescence immunoassay (ECLIA) (Elecsys, Anti-SARS-CoV-2, Roche Diagnostics) in sera of all participants. Results: Between February 2021 and April 2021, study questionnaires were completed for 253 pwMS (median age 47 years, 162 female). 211 pwMS (83%) had a relapsing, 25 (10%) a secondary progressive, 13 (5%) a primary progressive disease course and 4 (2%) a clinically isolated syndrome. Median disease duration was 12 years and median EDSS was 2.5. 218 (86%) pwMS were treated with DMTs: Ocrelizumab (27%), fingolimod (26%), dimethyl fumarate (15%), rituximab (10%), natalizumab (9%), other DMTs (13%). 15 (5.9%) of 253 pwMS had a positive SARSCoV- 2 PCR test since March 2020. In these pwMS, COVID-19 severity ranged from 1-10 on the WHO clinical progression scale: 1 pwMS was asymptomatic, 10 pwMS were symptomatic as outpatients (8 independently, 2 needed assistance), 3 pwMS were hospitalized (1 without oxygen therapy, 2 with oxygen by mask or nasal prongs) and 1 pwMS died. By April 2021, 24 and 38 pwMS received one and two doses of SARS-CoV-2 mRNA vaccines, respectively. Conclusions and outlook: Since start of the pandemic, rate of PCR-confirmed SARS-CoV-2 infection in our sample was slightly lower compared to incidence of laboratory-confirmed cases in Switzerland. The majority of pwMS had mild COVID-19. The study will continue until 2024 and by ECTRIMS 2021, we anticipate a doubling of completed questionnaires and to report preliminary results of serological measurements. This will allow us to present vaccine- and natural infection induced serological anti-SARS-CoV-2 responses in pwMS and assess differences related to various DMTs or COVID-19 severity.
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Background: The global SARS-CoV-2 outbreak has significantly affected hospital assistance to cancer patients. Diagnostic and treatment paradigms have been challenged with an urgent need for patient protection. In the abscence of data to balance clinical decissions we aimed to analyze HUIS experience during the peak of the outbreak. Method(s): Cancer pts atended at HUIS since February 24th to April 24th were collected. Clinical management was adapted according to evolving international consensus. All PCR+ COVID-19 pts have been included in a database. Oncological and COVID-19 diseases characteristics as well as cancer management have been collected. The main objective of this analysis was to know the risk of SARS-CoV-2 infection, hospitalization rate and mortality of cancer patientes in our center during the outbreak and to identify potential predictive factors. Result(s): Overall, 853 cancer pts had been attended at our department during this period of time. Twenty-six pts (3.05%) were hospitalized with confirmed COVID-19 diagnosis. Underlying solid tumors were the following: breast (256, 30.01%), GI (312, 36.8%), lung (100, 11.72%) and others (185, 21.47%). 322 pts (37.75%) had metastatic cancer and 531 (62.25%) had early stage diseases. 395 pts (46.31%) were treated with antineoplastic agents: 62,63% received treatment as adjuvant therapy, 18. 92% as first line (or maintenance) treatment in advanced diseases and 12.81% second or following lines of treatment in advaced diseases. 10 pts (32.26%) with COVID-19 died. Futher analysis regarding clinical, laboratory and hospital risk factors - such as diagnostic procedures, type and length of treatments, number of hospital visits, etc will be reported in the final presentation. Conclusion(s): COVID-19 hospitalization rate was 3.05%, and mortality rate was 32.26%. Adequate testing and protective measures are mandatory to warrant an optimal managment of cancer pts during the global SARS-CoV-2 outbreak. Legal entity responsible for the study: The authors. Funding(s): Has not received any funding. Disclosure: All authors have declared no conflicts of interest. Copyright © 2020