Effectiveness of ACB-IP 1.0 Universal Pathogen Free Concentrated Cocktail Convalescent Plasma in COVID-19 Infection

doi:10.21203/rs.3.rs-1042702/v1

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Effectiveness of ACB-IP 1.0 Universal Pathogen Free Concentrated Cocktail Convalescent Plasma in COVID-19 Infection

https://doi.org/10.21203/rs.3.rs-1042702/v1

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BACKGROUND

The efficacy of SARS-CoV2 single donor convalescent plasma (CP) varied according to the application time and the amount of antibody that is administered. Single donor CP has some drawbacks; such as the insufficient levels of neutralizing antibody activities, the requirements of blood group compatibility, and the risk of infection transmission. In this study, the safety and efficacy of pathogen inactivated, isohemagglutinin-depleted (concentrated) and pooled CP product was investigated.

MATERIALS AND METHODS

A total of sixteen patients were treated with either single donor CP (N:9) or pathogen-free, concentrated, pooled CP (ACB-IP 1.0) (N:7).

RESULTS

Five out of six single donor plasma SARS-CoV2 antibody titers remained below 12 s/co, but the antibody titers of all ACB-IP 1.0 plasma were above 12 s/co. SARS-CoV2 total antibody titers of ACB-IP 1.0 plasma were statistically higher than the antibody titers of single donor CP. Mean total plasma neutralizing antibody activity of ACB-IP 1.0 plasma (1.5421) was found statistically higher than single donor CP (0.9642) in 1:256 dilution (ρ<0.01)

The mortality rates of the patients treated with ACB-IP 1.0 plasma were statistically lower (p< 0.05) than the patients treated with single donor CP. The administration of either single donor CP or ACB-IP 1.0 plasma to the patients within eight days significantly shortened the length of hospitalization (ρ< 0.01).

CONCLUSION

The present study established ACB-IP 1.0 plasma product as a safe and potentially effective treatment for COVID-19, allowing rapid access to patients in need.

TRIAL REGISTRATION

Trial Registration Number: NCT04769245

Trial Registration Date: 17.03.2021

General Microbiology

Infectious Diseases

convalescent plasma

SARS-CoV2

COVID-19

neutralizing antibody

On 31 December 2019, a new case of pneumonia of anonymous etiology emerged in Wuhan City, Hubei Province China and humanity was confronted with a new pandemia. The World Health Organization (WHO) officially announced the causative organism as 2019-nCoV/SARS-CoV2.(1) Viral genome sequence of this new human pathogen was released and found to be closely related to viral species called Severe Acute Respiratory Syndrome Coronavirus (SARS-CoV) which caused outbreaks in 2002 and 2003 in China.(2) The SARS-CoV2 virus has spread from Wuhan to whole of China and 223 countries worldwide, affecting more than 208 million individuals and resulting over 4,300,000 million deaths.(3)

WHO recommends the use of Ivermectin, thromboprophylaxis, IL-6 blocker, Lopinavir/Lipanavir, Colchicine, Vitamin D, and corticosteroid in treatment of COVID-19.(4) Nevertheless, the exact clinical benefit of these COVID-19 therapeutics are not yet determined. Hence, clinicians seek for alternative treatment options such as monoclonal antibodies and convalescent plasma. Fundamental mechanism behind monoclonal antibodies and convalescent plasma are similar with both therapies generating passive immunization for viral neutralization.(5) However, convalescent plasma may be preferable over monoclonal antibodies in terms of being effective against new strains, being accessible and cost-effective. Furthermore, convalescent plasma is the safest treatment alternative for emergency use. Convalescent plasma containing antibody from SARS-CoV2 individuals who have recovered from the disease, has been suggested as an investigational treatment option by FDA.(6)

Our knowledge regarding convalescent human plasma comes from the treatment of viral infections, such as SARS-CoV, avian influenza A (H5N1) virus, influenza A (H1N1), MERS, and Ebola virus.(7-13) Even though there is still no consensus about its effectiveness, efficacy of convalescent plasma varied according to virus type, time of application, amount of the antibody and most importantly to the neutralizing capacity of the antibodies administered.(14) There are similar concerns regarding studies conducted in SARS-CoV2. FDA report and Mayo Clinic study involving 20,000 hospitalized patients indicated that early and adequate plasma use would reduce mortality(15, 16), while PLACID and the latest published RECOVERY trial reported single donor convalescent plasma did not improve survival or other clinical outcomes.(17, 18)

Conflicting outcomes of these studies may be due to the different times of administration, donor antibody titers (since SARS-CoV2 antibody titer was perceived to be the same as neutralizing antibody titer and the latter was therefore not estimated in most studies) and whether the convalescent plasma is strain-specific. Variations in these parameters have made the results debatable.

Single donor plasma has some drawbacks such as requirements of blood group compatibility(17, 19, 20) and risk of viral infection, for instance HIV, HBV, HCV transmission by donors who do not meet the standard donor criteria in pandemia conditions. In addition, excess of procoagulant factors in standard donor plasma increases the risk of thrombosis.(21, 22) These factors necessitated the search for a new convalescent plasma product. Hence, ACB-IP 1.0 cocktail plasma, where SARS-CoV2 antibody titers and neutralizing antibody activity were standardized by pooling, was designed. Furthermore, Immunoglobin M (IgM) which is responsible for most of the isohemagglutinins(23-25), was depleted by using both cryodepletion(24) and pooling method until Anti-A and Anti-B isohemagglutinin titers were below 1/8. Procoagulant factors were depleted by cryodepletion in order to eliminate the risk of thrombotic events. Furthermore, it was concentrated and safety of the plasma was improved by subjecting it to pathogen inactivation.

This study was conducted with participation of 16 patients, and product quality and clinical effectiveness of ACB IP 1.0 with standard single donor convalescent plasma were compared.

According to the criteria specified in the COVID-19 Immune Plasma Procurement and Clinical Use Guidelines of FDA(8), donor candidates who were eligible according to apheresis donor criteria were invited to Therapeutic Apheresis Center of Acıbadem Altunizade Hospital. Blood products were taken from the donor plasma candidates. (Supplement-1)

Effect of administration of convalescent plasma samples obtained from 9 Turkish Red Crescent donors and 7 ACB-IP 1.0 products, where each one of the 7 plasma samples was prepared from 8 different donors, were compared according to outcome of the patients.

Plasma Collection:

ACB IP 1.0 was obtained from donor plasma using Trima Accel (Trima Accel, Terumo BCT, Inc. Colorado, USA) device. 400 ml - 600 ml plasma was collected according to the patient’s height, weight, and blood count results. During this process, an ISBT code was obtained from the Turkish Red Crescent.

Pathogen Inactivation:

Plasma collected by plasmapheresis was connected to Cerus Intercept Blood System plasma treatment bags (Intercept Blood System Pathogen Reduction System, Cerus, CA, USA) using bag joining device (Terumo TSCD-II TSCD Welders, Terumo BCT, Inc. Colorado, USA). Prior to pathogen inactivation process, 2 ml of 2 tubes of witness samples were removed from the collected plasma. Witness samples were stored at -40 °C. According to the manufacturer instruction, plasma was initially treated with Amotosalen followed by photochemical irradiation with UVA at 320-400 nm wavelengths in Intercept INT100 illuminator (INTERCEPT INT100 Illuminator, Cerus, CA, USA). Following inactivation, plasma was passed through the adsorption filter to remove unreactive amotosalen and free photoproducts, and then it was divided into 2 or 3 equal volumes, depending on the volume of the collected plasma. Following this process, 2 ml of 2 witness sample tubes were separated and stored at -40 °C. Pathogen inactivated plasma was stored at -40 °C for labeling until the pooling process prior to clinical usage.

Isohemagglutinin Assay:

Ready to use (commercial) A and B kits (ID-Diacell ABO, Bio-Rad Laboratories, Inc., Cressier, Switzerland) were provided by Bio-Rad and gel centrifugation method was performed according to the manufacturer's protocol (NaCl, Enzyme Test and Cold Agglutinins, Bio-Rad Laboratories, Inc., Cressier, Switzerland).(26)

IgM Detection:

IgM titers were obtained by using Siemens Advia 1800 Chemistry System. The photometric method was performed according to the manufacturer’s protocol.

Isohemagglutinin Depletion and Concentration of Plasma:

Following obtaining plasma from the donor, Anti-A and Anti-B hemagglutinin titers were determined. Management of Isohemagglutinin titer was carried out in two separate steps. Firstly, isohemagglutinins, most of which were of IgM nature, were reduced by cryodepletion, while simultaneously concentrating the product. Secondly, isohemagglutinin titer was reduced by pooling of Anti- A and Anti-B free plasma and the plasma containing them.

Cryodepletion Method(23, 24)

Plasma samples from the apheresis product of 200 ml were transferred to plasma storage bags (Teruflex, Terumo BCT, Inc. Colorado, USA) frozen in a deep freezer at -40 °C. One bag consisted of eight donors’ apheresis plasma and volume in each bag was approximately 1600 ml. Frozen samples were kept at + 4 °C for defrosting for approximately 8-12 hours. Liquid plasma was separated from cryoprecipitate by centrifugation. Witness samples were taken and stored at -40 °C. The bag where cryodepleted pool would be produced was connected to the plasma bag (Terumo BCT, Inc. Colorado, USA) using the bag joining device (Terumo TSCD-II TSCD Welders, Terumo BCT, Inc. Colorado, USA).

Plasma Pooling:

Plasma bag was placed in the extractor and the cryopoor plasma was transferred to the pooling bag. Finally, pooling was achieved by mixing the low SARS-CoV2 antibody titer with high antibody titer plasma prior to cryodepletion and the total product was packaged in 200 ml bags and stored at -40 °C until clinical use.

SARS- CoV2 Specific Immunoglobulin Analysis:

SARS-CoV2 RBD specific total antibody analyzes were performed using CLIA method ( ADVIA Centaure XP, Siemens Healthineers, Erlangen, Germany).

Neutralizing Antibody Assay:(27)

Modified micro neutralization test was performed. 100 TCID50 / 50 microliter SARS-CoV2 virus was placed in 96 Well U Bottom plate and 50 µl diluted human sera ( 1:64, 1:128, 1:256 sera concentration) were added.

Following one hour of incubation at room temperature, 10000 Vero cell/well was placed in a 96 Well Flat Bottom plate with 100 µl of complete DMEM (4% FBS + 1% PSA). Supernatants were removed after 72 hours of incubation, 50 μl of MTT solution and 50 μl serum-free media were added to the remaining cells. Following incubation at 37 ° C for 4 hours, 100 μl of Isopropanol dispersion was added to each well and placed on a shaker for 10 min. Results were obtained by ELISA Reader at an absorbent value of 570 nm. Neutralizing antibody activity was studied in 1:64, 1:128 and 1:256 dilution based by cell viability index.

Endotoxin Analysis:

Gel-clot technique was used for detecting or quantifying endotoxins (Gel-Clot Endotoxin Test, Division of Charles River Laboratories Inc, CA).

Microbiological Quality Control:

Pooled convalescent plasma was placed on to Bactec Fx device for microbiological quality control analysis (Bactec FX, Becton Dickinson, New Jersey, USA).

Sars-CoV2 Quantitative Real-Time Polymerase Chain Reaction (PCR) Test:(28)

Nasopharyngeal swap sample was obtained by an experienced healthcare provider from a COVID-19 positive patient for the qualitative detection of nucleic acid from SARS-CoV2 in upper respiratory specimens. Analysis was performed by using Quantitative Real-Time PCR Coronavirus Detection test kit according to the manufacturer’s instructions (QuantiVirus™ SARS-CoV-2 Test kit, Diacarta, CA, USA)

Trial Design (Trial is recorded under; NCT04769245):

A total of 16 hospitalized adults were screened for enrollment and included in the study if they had positive reverse-transcriptase–polymerase-chain-reaction (RT-PCR) for SARS-CoV2 and radiologically confirmed pneumonia.

A total of 16 patients were treated with two different convalescent plasma products. Nine patients were treated with single donor convalescent plasma and seven were treated with pathogen-free, concentrated, pooled convalescent plasma between 01 March 2020 and 31 December 2020.

Written informed consent was obtained from all patients or their first degree relatives, and trial was conducted under the principles stated in the Declaration of Helsinki and Good Clinical Practice guidelines and approval of Acıbadem University ethics committee (Approval No: 2020-06/02).

Clinical information of the patients was obtained from the hospital’s electronic medical records. Demographic data, presenting symptoms as well as a radiological presentation at the onset of disease including fever, cough, fatigue, dyspnea, diarrhea, oxygen requirement, treatments received, duration of hospitalization stay, duration of Intensive care unit (ICU) stay, cycles and volume of convalescent plasma received, symptom and radiological improvements and current status of the patients were collected. Patients in the two groups were compared according to safety and efficacy, and followed up for transfusion-related reactions and findings recorded.

Statistical Analysis:

SARS-CoV2 Antibody titers, neutralizing antibody activities, and duration of hospitalization were analyzed by employing Mann Whitney test. Mean age of the groups was compared by the Kolmogorov-Smirnov test. Survival differences between the two plasma groups were analyzed by Chi-Square test. Fisher exact test was used to compare the categorical variables such as radiological presentation, co-existing disease and oxygen supplement requirement among the groups. Results were analyzed with a %95 confidence interval and a significance level of p=0.05 was used for all statistical analysis.

Laboratory Results:

Results of the Isohemagglutinin Titers:

Pre-pooling cryodepletion process caused a statistically significant reduction of 35% in Anti-A titers, 44% in Anti-B titers, and 11% in IgM levels (Table 1, Figure 1). Following the pooling process, maximum Anti-A titers in ACB-IP 1.0 was reduced to 1/4 and Anti-B titers to 1/8, while maximum Anti-A titers were 1/64 and Anti-B 1/128 in single donor plasma. Results show that cryodepletion and pooling process effectively control isohemagglutinin levels, giving ACB-IP 1.0 a universal character (blood-group free convalescent plasma).

Results of the SARS-CoV2 Antibody Titers in Plasma:

Five out of six single donor plasma SARS-CoV2 antibody titers remained below 12 s/co, but antibody titers of all ACB-IP 1.0 plasma were above 12 s/co. Mean antibody titer of single donor plasma was 9.2083 and mean antibody titer of ACB-IP 1.0 plasma was 32.700. SARS-CoV2 total antibody titers of ACB-IP 1.0 plasma were statistically higher in comparison with the antibody titers of single donor plasma (Figure 2).

Results of SARS-CoV2 Neutralizing Antibody Activity:

SARS-CoV2 neutralizing capacity of single donor plasma antibodies was higher than SARS-CoV2 antibody titers. Only 50% single donor plasma neutralizing antibody activity was below %100 cell viability threshold in 1:256 dilution (Figure 3), whereas neutralizing activity of all ACB-IP1.0 plasma was above %100 cell viability threshold in 1:256 dilutions (Figure 3).

Mean total plasma neutralizing antibody activity of ACB-IP 1.0 plasma (1.5421) was found to be statistically higher than single donor plasma (0.9642) in 1:256 dilution (ρ=**0.0087) (Figure 3).

No correlation was found between SARS-CoV2 antibody titers and neutralizing antibody activity in both groups (single donor plasma antibody and neutralizing antibody activity correlation coefficient: 0.54, regression coefficient (R²): 0.44. ACB-IP 1.0 plasma antibody and neutralizing antibody activity correlation coefficient: -0.36, regression coefficient (R²): 0.79).

Clinical Results:

Clinical Characteristics of Single Donor plasma patients were presented in Table-2. Six out of nine single donor convalescent plasma patients were male. Median age of the patients was 65 years and none of them had a history of smoking. Two male patients aged 54 and 63 had no coexisting disease.

Treatment of all patients was performed according Covid-19 treatment algorithms. Five patients received dornase-alpha and three patients received IL-6 blocker.

One of the patients had two cycles of single donor convalescent plasma total of 400 ml volume.
Volume loading due to transfusion was detected in one patient. No other reaction was observed.

Mean duration of hospital stay was 41 days (11-101 days) and mean duration of ICU stay was 35 days (7-90 days). Following the treatments and convalescent plasma administration, four out of nine patients had clinical improvement, three patients had radiological improvement, one of the patients was not evaluated. Five out of nine patients died in total.

Clinical characteristics of SARS-CoV2 infected patients who received pathogen-free, concentrated, pooled, convalescent plasma is presented in Table-3. All of the patients treated with pathogen-free, concentrated, pooled convalescent plasma were males. Median age of the patients was 51 years and none of them had a history of smoking. Two patients aged 50 and 58 had no coexisting disease.

Treatments of all seven patients were performed according to Covid-19 treatment algorithms. One patient received IL-6 blocker and one patient had dornase-alpha treatment.

One of the patients had two cycles of pooled convalescent plasma total of 400 ml volume. No transfusion-related reactions had been observed. The mean duration of the hospital stay was 44 days (12-172 days) and the mean duration of ICU stay was 30 days (0-139 days).
Following treatments and convalescent plasma administration, four patients had clinical improvement and five patients had radiological improvement. Seven patients in total were discharged from the hospital.

There was no statistical difference between groups of patients that received single donor plasma and ACB-IP 1.0 plasma with respect to age (ρ=0.1604), radiological presentation (ρ= 0.999), co-existing disease (ρ= 0.999) and oxygen supplement requirement (ρ= 0.5962).

Mortality rate of the patients treated with ACB-IP 1.0 plasma were statistically lower (p: 0.033) than the patients treated with single donor plasma. (Figure 4)

Median length of hospital stay was 41 days for single donor plasma patients, and 44 days for ACB-IP 1.0 plasma patients and there was no significant difference. Administration of single donor plasma or ACB-IP 1.0 plasma to the patients within eight days from presentation significantly shortened the length of hospitalization in comparison with administration of either plasma later than eight days from presentation (ρ= 0.0021) (Figure 5).

There are conflicting results regarding the beneficial effect of administering convalescent plasma in SARS-CoV2 pneumonia. An initial report of five critically ill patients with COVID-19 and Acute Respiratory Distress Syndrome (ARDS), where single dose of convalescent plasma was administered, indicated clinical improvement.(24) In another study, convalescent plasma of 200ml was administered to ten patients with COVID-19 infection. Results showed that patients had clinical benefit, and common laboratory parameters of infection such as decreased lymphocyte count and increased CRP normalized.(19) A recent prospective and propensity score-matched study, comparing survival rates of convalescent plasma transfused 136 patients with non-transfused 251 patients, recommended administration of convalescent plasma within 72 hours of hospital admission due to observed significant reduction in mortality rate.(29) In a matched study, 20 severely and critically ill hospitalized COVID-19 patients and 20 controls were examined according to their laboratory, respiratory parameters and mortality rate. Although the study had some limitations, such as short follow-up time and small sample size, results showed that convalescent plasma could improve survival if given at the early onset of disease.(17) Liu et al. reported 39 hospitalized patients with severe life-threatening COVID-19, who received convalescent plasma, compared with a non-transfused control group. Results showed that patients receiving convalescent plasma therapy had improved survival and reduced oxygen requirement 14 days post-transfusion. In this study, authors also recommended transfusion of convalescent plasma immediately following hospitalization.(30) Mayo Clinic study involving 20,000 hospitalized patients and a randomized controlled study conducted by Libster et al. in elderly patients also suggested that early administration of convalescent plasma reduced clinical symptoms and progression of Covid-19 to severe illness.(16, 31)

On the other hand, a number of studies failed to show beneficial effect of convalescent plasma in SARS-CoV2 pneumonia patients.(17, 18, 32) One of these studies was PLACID trial, even though it had a number of design faults, such that there was no neutralizing antibody titer measurement of the donor plasma and also convalescent plasma was administered to the patients later than 3 days, contrary to what was recommended by the FDA. V.A. Simonovich et al. also showed no relationship between SARS-CoV2 antibody titers, early plasma administration and clinical efficacy.(32) However, hypoxia was one of the patient selection criteria and treatment was not commenced within 72 hours of the disease onset and neutralizing antibody titers could only be assessed in 56% of the donors in this study.

Largest trial conducted so far, RECOVERY Group trial, also indicated that even if convalescent plasma is used in early onset of disease, it does not reduce respiratory support requirement, hospital stay, and mortality.(18) However; convalescent plasma in this study was again administered later than 72 hours following the onset of disease. Also the convalescent plasma was not strain-specific because a new SARS- CoV-2, B.1.1.7 dominated most regions of UK at the time of the study. Hence, it is likely that modified antigenicity further reduced neutralizing capacity of the plasma. Furthermore, all convalescent plasma were chosen based on high anti-spike concentration. This was on the assumption that IgG concentration shows correlation with the neutralizing antibody titer.(18) Although, Salazar et al. showed that there is a correlation between antibody titers and neutralizing capacity, they only reported this correlation in only 80% of single donor plasma. The fact that the correlation between IgG concentration and the neutralizing antibody titer was not shown in %20 of single donor plasma is a high ratio that can make the results questionable.(33)

Results of the current study for single donor plasma showed heterogeneity of SARS-CoV2 antibody titers and neutralizing antibody activity. However, ACB-IP 1.0 plasma which is a pooled concentrated plasma product, showed higher SARS-CoV2 antibody titers and neutralizing antibody activity. There was no correlation between SARS-CoV2 antibody titers and neutralizing antibody activities in single donor plasma or ACB-IP 1.0 plasma. This may have been due to only SARS-CoV-2 S1 spike protein antigen being used for antibody detection in this study. More reliable results could possibly have been obtained using nucleocapsid and matrix protein antigen in addition to the spike protein antigen for antibody detection.

Presence of re-infected cases despite high antibody titers confirms that this is not a reliable parameter for protection against Covid. ACB-IP 1.0 plasma in this study had less variance of SARS-CoV2 antibody and neutralizing antibody titer, and hence more standardized convalescent plasma could be obtained.

Findings of this study showed significantly decreased in mortality in ACB-IP 1.0 plasma in comparison with single donor convalescent plasma. There was also a reduction of length of hospital stay in both groups if the plasma was administrated within 8 days onset of disease (p=0.0021). Failure to show significant difference at 3 days may have been due to the small number of patients in this study group.

Another, significant clinical result was difficulty in accessibility of single donor plasma, which requires blood group compatibility, especially in the early stages of the pandemic due to the lack of sufficient available donors. However, as a result of cryodepletion and pooling processes, ACB-IP 1.0 convalescent plasma does not require blood group compatibility, and hence can be employed immediately. Furthermore, by containing pooling of plasma from various convalescent individuals, ACB-IP 1.0 plasma may be more advantageous in the treatment of variant mutations.

Single donor plasma also has the disadvantage of increased risk of transfusion-transmitted infections due to individuals who may not meet the standard donor criteria. With the help of pathogen inactivation process, transfusion-transmitted infections are minimized. However, due to its small scale, no data could be obtained to determine the advantage of pathogen inactivation in this study and a randomized, controlled study with a larger patient group is required.

ACB-IP 1.0 pooled pathogen inactive universal convalescent plasma leads to a better clinical response in comparison with single donor CP with a neutralizing antibody activity against SARS-CoV2. The product is safe and does not require blood group compatibility allowing fast access for patients in need.

ARDS: Acute Respiratory Distress Syndrome

CP: Convalescent plasma

CRP: C-Reactive Protein

FDA: Food and Drug Administration

HBV: Hepatitis B Virus

HCV: Hepatitis C virus

HIV: Human Immunodeficiency Virus

H1N1: Influenza A virus subtype

H5N1: Influenza A virus subtype

IGG: Immunoglobulin G

IGM: Immunoglobulin M

IL-6: Interleukin-6

MERS: Middle East respiratory syndrome

RT-PCR: Reverse transcriptase polymerase chain reaction

SARS-CoV2: Severe acute respiratory syndrome coronavirus 2

s/co: Signal-to-cutoff

UVA: long wave ultraviolet A

WHO: World Health Organization

Ethics approval and consent to participate: Acıbadem University Ethics Committee (Approval No: 2020-06/02).

Consent for publication: Not Applicable

Availability of data and materials: All data generated or analyzed during this study are included in this published article

Competing interests: The authors declare that they have no competing interests

Funding: All funding of the work was supported by TUBITAK

Authors' contributions:Data Collection or Processing: C.H.,O.S.G., N.O.S., E.S.K., O.E., S.O.D., E.S., R.D.T., G.C., M.E.K., G.S.K., B.Y., C.T., D.D.K., Z.T.,U.T.,M.E.,R.Z., A.S.K, C.C., N.B. Analysis or Interpretation: .C.H., N.B.P.,K.Y., R.D.T., S.A., D.C., E.O. Writing: C.H., K.Y., S.R., E.O.

Acknowledgements: Not Applicable

(WHO) WHO. Coronavirus Geneva WHO 2020 https://www.who.int/health-topics/coronavirus Access Date 21.1.2021. 2020.
Peiris JS, Yuen KY, Osterhaus AD, Stohr K. The severe acute respiratory syndrome. The New England journal of medicine. 2003;349(25):2431-41. Epub 2003/12/19.
https://www.worldometers.info/coronavirus/ 2021.
https://www.who.int/publications/i/item/WHO-2019-nCoV-therapeutics-2021.1. 2021.
Casadevall A, Pirofski LA. The convalescent sera option for containing COVID-19. The Journal of clinical investigation. 2020;130(4):1545-8. Epub 2020/03/14.
DM. H. Emergency use authorization for COVID-19 convalescent plasma therapy. US Food and Drug Administration website. Published August 24, 2020. Accessed January 21, 2021. https://www.fda.gov/media/141477/download. 2020.
Who Mers-Cov Research G. State of Knowledge and Data Gaps of Middle East Respiratory Syndrome Coronavirus (MERS-CoV) in Humans. PLoS currents. 2013;5:ecurrents.outbreaks.0bf719e352e7478f8ad85fa30127ddb8.
Hung IFN, To KKW, Lee CK, Lee KL, Yan WW, Chan K, et al. Hyperimmune IV immunoglobulin treatment: a multicenter double-blind randomized controlled trial for patients with severe 2009 influenza A(H1N1) infection. Chest. 2013;144(2):464-73. Epub 2013/03/02.
Stockman LJ, Bellamy R, Garner P. SARS: systematic review of treatment effects. PLoS medicine. 2006;3(9):e343. Epub 2006/09/14.
Hung IF, To KK, Lee CK, Lee KL, Chan K, Yan WW, et al. Convalescent plasma treatment reduced mortality in patients with severe pandemic influenza A (H1N1) 2009 virus infection. Clinical infectious diseases : an official publication of the Infectious Diseases Society of America. 2011;52(4):447-56. Epub 2011/01/21.
Luke TC, Kilbane EM, Jackson JL, Hoffman SL. Meta-analysis: convalescent blood products for Spanish influenza pneumonia: a future H5N1 treatment? Annals of internal medicine. 2006;145(8):599-609. Epub 2006/08/31.
Cheng Y, Wong R, Soo YO, Wong WS, Lee CK, Ng MH, et al. Use of convalescent plasma therapy in SARS patients in Hong Kong. European journal of clinical microbiology & infectious diseases : official publication of the European Society of Clinical Microbiology. 2005;24(1):44-6. Epub 2004/12/24.
Ko JH, Seok H, Cho SY, Ha YE, Baek JY, Kim SH, et al. Challenges of convalescent plasma infusion therapy in Middle East respiratory coronavirus infection: a single centre experience. Antiviral therapy. 2018;23(7):617-22. Epub 2018/06/21.
Wang Y, Huo P, Dai R, Lv X, Yuan S, Zhang Y, et al. Convalescent plasma may be a possible treatment for COVID-19: A systematic review. International immunopharmacology. 2021;91(107262):5.
Joyner MJ, Senefeld JW, Klassen SA, Mills JR, Johnson PW, Theel ES, et al. Effect of Convalescent Plasma on Mortality among Hospitalized Patients with COVID-19: Initial Three-Month Experience. medRxiv : the preprint server for health sciences. 2020:2020.08.12.20169359.
Joyner MJ, Bruno KA, Klassen SA, Kunze KL, Johnson PW, Lesser ER, et al. Safety Update: COVID-19 Convalescent Plasma in 20,000 Hospitalized Patients. Mayo Clinic proceedings. 2020;95(9):1888-97.
Agarwal A, Mukherjee A, Kumar G, Chatterjee P, Bhatnagar T, Malhotra P. Convalescent plasma in the management of moderate covid-19 in adults in India: open label phase II multicentre randomised controlled trial (PLACID Trial). BMJ. 2020;22(371).
Convalescent plasma in patients admitted to hospital with COVID-19 (RECOVERY): a randomised controlled, open-label, platform trial. Lancet. 2021;397(10289):2049-59.
Duan K, Liu B, Li C, Zhang H, Yu T, Qu J, et al. Effectiveness of convalescent plasma therapy in severe COVID-19 patients. Proceedings of the National Academy of Sciences of the United States of America. 2020;117(17):9490-6.
Shen C, Wang Z, Zhao F, Yang Y, Li J, Yuan J, et al. Treatment of 5 Critically Ill Patients With COVID-19 With Convalescent Plasma. Jama. 2020;323(16):1582-9.
Zander AL, Olson EJ, Van Gent JM, Bandle J, Calvo RY, Shackford SR, et al. Does resuscitation with plasma increase the risk of venous thromboembolism? The journal of trauma and acute care surgery. 2015;78(1):39-43.
Holcomb JB, del Junco DJ, Fox EE, Wade CE, Cohen MJ, Schreiber MA, et al. The prospective, observational, multicenter, major trauma transfusion (PROMMTT) study: comparative effectiveness of a time-varying treatment with competing risks. JAMA Surg. 2013;148(2):127-36.
Hadjesfandiari N, Levin E, Serrano K, Yi Q-L, Devine DV. Risk analysis of transfusion of cryoprecipitate without consideration of ABO group. Transfusion. 2021;61(1):29-34.
Meliga SC, Farrugia W, Ramsland PA, Falconer RJ. Cold-Induced Precipitation of a Monoclonal IgM: A Negative Activation Enthalpy Reaction. The Journal of Physical Chemistry B. 2013;117(2):490-4.
Middaugh CR, Gerber-Jenson B, Hurvitz A, Paluszek A, Scheffel C, Litman GW. Physicochemical characterization of six monoclonal cryoimmunoglobulins: possible basis for cold-dependent insolubility. Proceedings of the National Academy of Sciences of the United States of America. 1978;75(7):3440-4. Epub 1978/07/01.
Josephson CD, Mullis NC, Van Demark C, Hillyer CD. Significant numbers of apheresis-derived group O platelet units have "high-titer" anti-A/A,B: implications for transfusion policy. Transfusion. 2004;44(6):805-8.
Nie J, Li Q, Wu J, Zhao C, Hao H, Liu H, et al. Quantification of SARS-CoV-2 neutralizing antibody by a pseudotyped virus-based assay. Nature protocols. 2020;15(11):3699-715.
Emery SL, Erdman DD, Bowen MD, Newton BR, Winchell JM, Meyer RF, et al. Real-time reverse transcription-polymerase chain reaction assay for SARS-associated coronavirus. Emerging infectious diseases. 2004;10(2):311-6.
Salazar E, Christensen PA, Graviss EA, Nguyen DT, Castillo B, Chen J, et al. Treatment of Coronavirus Disease 2019 Patients with Convalescent Plasma Reveals a Signal of Significantly Decreased Mortality. The American journal of pathology. 2020;190(11):2290-303.
Liu STH, Lin H-M, Baine I, Wajnberg A, Gumprecht JP, Rahman F, et al. Convalescent plasma treatment of severe COVID-19: a propensity score–matched control study. Nature medicine. 2020;26(11):1708-13.
Libster R, Pérez Marc G, Wappner D, Coviello S, Bianchi A, Braem V, et al. Early High-Titer Plasma Therapy to Prevent Severe Covid-19 in Older Adults. The New England journal of medicine. 2021;384(7):610-8.
Simonovich VA, Burgos Pratx LD, Scibona P, Beruto MV, Vallone MG, Vázquez C, et al. A Randomized Trial of Convalescent Plasma in Covid-19 Severe Pneumonia. New England Journal of Medicine. 2020;384(7):619-29.
Salazar E, Kuchipudi SV, Christensen PA, Eagar T, Yi X, Zhao P, et al. Convalescent plasma anti-SARS-CoV-2 spike protein ectodomain and receptor-binding domain IgG correlate with virus neutralization. The Journal of clinical investigation. 2020;130(12):6728-38.

Table 1: Effects of Cryodepletion on Isohemagglutinin and IgM levels of single donor plasma

N=8	Mean Anti-A titers	Mean Anti-B titers	Mean IgM titers
Prior to Cryodepletion	1/69,3±50,26	1/96,00±35,05	76,0±28,72
Following Cryodepletion	1/32,0±24,79^*	1/53,3±16,52^*	67,2±26,54^*
% Change	-53,85	-44,44	-11,62

* significant level by t test for p<0.05.

Table 2: Clinical Characteristics of SARS-CoV-2-Infected Patients Who Received Single Donor Convalescent Plasma Table-2.

Patient No	1	2	3	4	5	6	7	8	9
Sex	F	M	M	M	M	F	M	F	M
Age	71	59	40	54	63	65	72	76	71
Coexisting Chronic Diseases	Chronic Myeloid Leukemia, Coronary Heart Disease	Hyperlipidemia, Hypertension, Gastritis, Morbid Obesity	No	No	No	Hypertension	Ischemic heart disease	Hypertension, Ischemic heart disease, Emphysema	Hypertension, Heart disease, Atrial flutter
Fever	No	No	Yes	Yes	Yes	No	Yes	Yes	Yes
Cough	No	Yes	Yes	Yes	Yes	No	Yes	No	Yes
Fatigue	Yes	Yes	Yes	Yes	Yes	No	Yes	Yes	Yes
Dyspnea	Yes	Yes	Yes	Yes	Yes	Yes	Yes	Yes	No
Diarrhea	No	Yes	No	No	No	No	No	No	No
Oxygen Supplement Requirement	Yes	Yes	Yes	Yes	No	Yes	Yes	Yes	No
Radiological Presentation	Multiple GGOs with consolidation	Multiple GGOs with consolidation	Multiple GGOs with consolidation	Multiple GGOs with consolidation, Bronchopneumonia	Multiple GGOs with consolidation, Partial Air Bronchogram	Multiple GGOs with consolidation, Atelectasis	Multiple GGOs with consolidation	Density increase in heterogeneous structure compatible with consolidation	Multiple GGOs with consolidation, Interstitial septal thickening, Air cyst, Band atelectasis
Length of stay In Hospital	24	11	31	101	23	20	98	23	38
Length of stay In ICU	24	7	21	90	21	20	73	23	35
Convalescent Plasma Application Date	2.May	24 April	11 April	19 April	17 April	19 April	27 April	9.May	23 April
Interval Between Disease On Set and Plasma Transfusion	11	6	15	26	18	16	35	5	28
Cycle of Convalescent Plasma	1	1	1	2	1	1	1	1	1
Convalescent Plasma Volume (ml)	200 m l	200 m l	200 ml	400 m l	200 ml	200 ml	200 m l	200 m l	200 m l
Complications Prior Plasma Transfusion	No	No	No	No	No	No	No	No	No
Symptom Improvement	No	Yes	Yes	Yes	No	No	Yes	No	N o
Radiological Improvement	Not evaluated	Yes	No	Yes	No	Very Little İmprovement	No	No	Not evaluated
Current Status	Exitus	Discharged home	Discharged home	Discharged home	Exitus	Exitus	Discharged home	Exitus	Exitus

Table 3: Clinical Characteristics of SARS-CoV-2-Infected Patients Who Received Pathogen-Free, Concentrated, Pooled, Convalescent Plasma Table-3.

Patient No	1	2	3	4	5	6	7
Sex	Male	Male	Male	Male	Male	Male	Male
Age	49	72	51	50	41	59	56
Coexisting Chronic Diseases	Hypertension	Hypertension, Chronic Obstructive Pulmonary Disease	Asthma, Hypertension, Psoriatic arthritis	No	Allergic Rhinitis	No	Coronary Artery Disease
Fever	Yes	No	No	Yes	Yes	Yes	No
Cough	Yes	Yes	No	Yes	Yes	No	Yes
Fatigue	No	Yes	No	Yes	Yes	Yes	Yes
Dyspnea	No	Yes	Yes	Yes	Yes	No	Yes
Diarrhea	No	Yes	No	No	No	Yes	No
Oxygen Supplement Requirement	No	Yes	Yes	Yes	No	No	Yes
Radiological Presentation	Multiple GGOs with consolidation	Multiple GGOs with consolidation	Multiple GGOs with consolidation and fibrosis streak - Patchy areas of GGOs	Multiple GGOs with consolidation	Multiple GGOs with consolidation and fibrosis streak	Multiple GGOs with consolidation	Multiple GGOs with consolidation and fibrosis streak
Length of stay In Hospital	12	172	13	19	24	5	61
Length of stay In ICU	No	139	No	10	24	No	37
Convalescent Plasma Application Date	13 April	16 April	7 April	23 April	2 May	15 December	20 July
Interval Between Disease On Set and Plasma Transfusion	1	1	1	1	2	1	1
Cycle of Convalescent Plasma	8	24	14	8	6	3	12
Convalescent Plasma Volume (ml)	144	156	170	136	142 & 144	200	176
Complications Prior Plasma Transfusion	No	No	No	No	No	No	No
Symptom Improvement	Yes	Yes	Yes	Yes	Yes	Yes	Yes
Radiological Improvement	Yes	Yes	No	No	Yes	Yes	Yes
Current Status	Discharged home	Discharged home	Discharged home	Discharged home	Discharged home	Discharged home	Discharged home

No competing interests reported.

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Effectiveness of ACB-IP 1.0 Universal Pathogen Free Concentrated Cocktail Convalescent Plasma in COVID-19 Infection

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