Comparative phenotypical analysis of B cells in fresh and cryopreserved mononuclear cells from blood and tissue of rhesus macaques

Although autoantibodies have been used for decades as diagnostic and prognostic markers in type 1 diabetes (T1D), further analysis of developmental abnormalities in B cells could reveal tolerance checkpoint defects that could improve individualized therapy. To evaluate B cell developmental progression in T1D, immunophenotyping was used to classify circulating B cells into transitional, mature naïve, mature activated, and resting memory subsets. Then each subset was analyzed for the expression of additional maturation-associated markers. While the frequencies of B cell subsets did not differ significantly between patients and controls, some T1D subjects exhibited reduced proportions of B cells that expressed transmembrane activator and CAML interactor (TACI) and Fas receptor (FasR). Furthermore, some T1D subjects had B cell subsets with lower frequencies of class switching. These results suggest circulating B cells exhibit variable maturation phenotypes in T1D. These phenotypic variations may correlate with differences in B cell selection in individual T1D patients.

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“…However, cryopreservation can induce significant changes in several critical B cell subsetting markers, including CD27 and CD38. [17]…”

Section: Introductionmentioning

confidence: 99%

Circulating B cells in type 1 diabetics exhibit fewer maturation-associated phenotypes

Hanley

Sutter

Goodman

et al. 2017

Clinical Immunology

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“…Cryopreservation has the potential to induce phenotypic changes and can drastically decrease cell viability. Changes induced by cryopreservation have been explored for Bcells [5][6][7][8] , T-cells 9,10 and other hematopoietic cell sub-populations 11,12 . Therefore, evaluating the differences between freshly isolated and frozen cells is necessary to understand the potential effects that cryopreservation may have on downstream functional analyses.…”

Section: Introductionmentioning

confidence: 99%

Novel Method Enabling the Use of Cryopreserved Primary Acute Myeloid Leukemia Cells in Functional Drug Screens

Degnin

Agarwal

Tarlock

et al. 2017

Journal of Pediatric Hematology/Oncology

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The ability to assess antileukemic drug activity on primary patient samples is a powerful tool in determining potential drug targets and selection of therapeutic agents with biological and functional rationale. We previously established small molecule inhibitor screens for use on freshly isolated leukemia cells for this purpose. Here we describe a method that produces functional small molecule inhibitor screening results using cryopreserved primary acute myeloid leukemia cells. This method was established to take advantage of biorepositories containing archival material, such as those established by the Children's Oncology Group, and to enable validation of potential pathway dependencies uncovered by genomic analysis. Various conditions used to thaw and culture cryopreserved specimens were assessed for effect on viability, differentiation, and the ability to recapitulate sensitivity results obtained on fresh samples. The most reproducible results were obtained by quick-thawing and culturing samples in cytokine rich media before performing drug screens. Our data suggest that cytokine-enriched media aids in maintaining the viability and numbers required to perform functional analysis on cryopreserved leukemia cells. This method can aid in producing informative data on therapeutic targeting and precision medicine efforts in leukemia by making use of biorepositories and bio banks.

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“…Another plausible reason for not detecting antigen‐specific ASCs in tonsils could be due to the fact that ELISpot for B cell detection was done on frozen MNCs. A study by Klippert et al in rhesus macaques showed significant differences in percentages of B cells in freshly isolated and cryopreserved MNCs in blood and different body organs.…”

mentioning

confidence: 99%

Detection of rotavirus‐ and norovirus‐specific IgG memory B cells in tonsils

Sharma

Hagbom

Nordgren

et al. 2018

Journal of Medical Virology

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Because rotavirus (RV) and norovirus (NoV) are transmitted through the fecal-oral route, tonsils due to their location within the oropharynx may sample or become infected with these viruses. We investigated if RV and NoV RNA/antigen, or virus-specific memory/plasma B cells can be detected in the tonsils. While neither RV/NoV antigen, nor genomic RNA was detected, 90% (27/30) of tonsils tested had RV- and NoV-specific IgG memory B cells. However, the mechanism explaining how these cells get there (whether because of local induction or homing after induction at other sites) and the role these cells might play during active infection is not yet clear.

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Comparative phenotypical analysis of B cells in fresh and cryopreserved mononuclear cells from blood and tissue of rhesus macaques

Cited by 5 publications

References 42 publications

Circulating B cells in type 1 diabetics exhibit fewer maturation-associated phenotypes

Circulating B cells in type 1 diabetics exhibit fewer maturation-associated phenotypes

Novel Method Enabling the Use of Cryopreserved Primary Acute Myeloid Leukemia Cells in Functional Drug Screens

Detection of rotavirus‐ and norovirus‐specific IgG memory B cells in tonsils

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