CD3+CD20+ cells may be an artifact of flow cytometry: Comment on the article by Wilk et al

Henry, Carole; Ramadan, Abdulraouf; Montcuquet, Nicolas; Pallandre, Jean-René; Mercier‐Letondal, Patricia; Deschamps, Marina; Tiberghien, Pierre; Ferrand, Christophe; Robinet, Éric

doi:10.1002/art.27527

Cited by 23 publications

(22 citation statements)

References 11 publications

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“…Subsequent confocal microscopy of these CD3 + /CD20 + /CD19 - cells (Figure 2C) demonstrated that they also express the T-cell marker CD4 or CD8. Importantly, CD3 + /CD20 + /CD19 - cells were clearly identifiable as single cells and not, as previously suggested, as T-cell/B-cell doublets [10]. Interestingly, the expression of CD20 on the T cells appears rather patchy and may reflect cross-linking of anti-CD20 to CD20, which has been observed to result in CD20 translocating to lipid rafts [13].…”

Section: Resultsmentioning

confidence: 73%

“…A criticism of the previous study by Wilk and colleagues [9] was that CD3 + CD20 + cells may be an artefact of T-cell selection and may actually represent T-cell/B-cell doublets [10]. Wilk and colleagues [31] vigorously rejected this possibility.…”

Section: Discussionmentioning

confidence: 99%

“…However, the extent and nature of this CD20 + T-cell subset in health and disease-and therefore the possible relevance of this subset in RA-are not yet known. In addition, some groups have suggested that these cells may be T-cell/B-cell doublets [10]. …”

Section: Introductionmentioning

confidence: 99%

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Frequency of Th17 CD20+ cells in the peripheral blood of rheumatoid arthritis patients is higher compared to healthy subjects

et al. 2011

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IntroductionRheumatoid arthritis (RA) is considered a T cell driven autoimmune disease, therefore, the ability of B cell depleting biologics, e.g., anti-CD20 antibodies, to alleviate RA is unclear. This study examined the proportions of IL-17-secreting lymphocytes in the blood of healthy subjects and RA patients and determined if Th17 cells belong to a CD20+ subset of T cells.MethodsFluorescence-activated cell sorting and confocal microscopy verified CD3, CD4/CD8 and CD20-staining of T cells. IL-17 secretion was determined using a commercial assay.ResultsIn healthy subjects and RA patients blood, the median percentage of total CD20+ lymphocytes was similar (7.5%; n = 6 and 10.3%; n = 9, respectively) and comprised predominantly of B cells (~ 86%). However, 2-4% of CD3+ T cells from both healthy subjects (n = 7) and RA (n = 8) individuals co-expressed CD20. The peripheral blood of healthy subjects contained few IL-17-secreting CD20+ T cells (< 0.1%; n = 6). In contrast, in RA blood a median and interquartile range % of, 24.2%; IQR 28.5 of IL-17-secreting T cells were CD20+ (n = 9; p = 0.02).ConclusionsIn the blood of RA patients, a greater proportion of Th17 cells are of a CD20+ phenotype compared to healthy individuals. These cells may represent an additional target for anti-CD20 therapies.

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Section: Resultsmentioning

confidence: 73%

Section: Discussionmentioning

confidence: 99%

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Frequency of Th17 CD20+ cells in the peripheral blood of rheumatoid arthritis patients is higher compared to healthy subjects

et al. 2011

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“…Low percentages of CD20-expressing T cells in human blood were first described in 1993 (6), but the existence of this rather rare T cell subset has been disputed (7). Others have found that CD20-expressing T cells can exhibit pro-inflammatory capacity (8, 9).…”

Section: Introductionmentioning

confidence: 99%

Rituximab Efficiently Depletes Increased CD20-Expressing T Cells in Multiple Sclerosis Patients

Palanichamy

Jahn

Nickles

et al. 2014

The Journal of Immunology

244

290

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In multiple sclerosis (MS4) B cell depleting therapy using monoclonal anti-CD20 antibodies, including rituximab (RTX) and ocrelizumab (OCR), effectively reduces disease activity. Based on indirect evidence, it is generally believed that elimination of the antigen presenting capabilities and antigen non-specific immune functions of B cells underlie the therapeutic efficacy. However, a small subset of T lymphocytes (T cells) was shown to also express CD20, but controversy prevails surrounding the true existence of this T cell subpopulation. Using single-cell imaging flow cytometry and expression profiling of sorted lymphocyte subsets, we unequivocally demonstrate the existence of CD3+CD20dim T cells. We show that in MS patients increased levels of CD3+CD20dim T cells are effectively depleted by RTX. The pathological relevance of this T cell subset in MS remains to be determined. However, given their potential pro-inflammatory functionality, depletion of CD20-expressing T cells may also contribute to the therapeutic effect of RTX and other monoclonal antibodies targeting CD20.

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“…The first report of CD20-expressing T cells in the circulation dates back to 1993, where a subset of CD3 + CD20 + T cells has been described in healthy subjects (7) and later also in peripheral T cell lymphoma (8). The description of CD3 + CD20 + T cells in rheumatoid arthritis patients and controls (9) was disputed and has been regarded as an artifact of flow cytometry (10). Yet recently, a subset of CD3 + CD20 + T cells that is also targeted by rituximab has been described (11).…”

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confidence: 99%

Features of Human CD3+CD20+ T Cells

Schuh

Berer

Mulazzani³

et al. 2016

The Journal of Immunology

159

200

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Monoclonal Abs against CD20 reduce the number of relapses in multiple sclerosis (MS); commonly this effect is solely attributed to depletion of B cells. Recently, however, a subset of CD3+CD20+ T cells has been described that is also targeted by the anti-CD20 mAb rituximab. Because the existence of cells coexpressing CD3 and CD20 is controversial and features of this subpopulation are poorly understood, we studied this issue in detail. In this study, we confirm that 3–5% of circulating human T cells display CD20 on their surface and transcribe both CD3 and CD20. We report that these CD3+CD20+ T cells pervade thymus, bone marrow, and secondary lymphatic organs. They are found in the cerebrospinal fluid even in the absence of inflammation; in the cerebrospinal fluid of MS patients they occur at a frequency similar to B cells. Phenotypically, these T cells are enriched in CD8+ and CD45RO+ memory cells and in CCR7− cells. Functionally, they show a higher frequency of IL-4–, IL-17–, IFN-γ–, and TNF-α–producing cells compared with T cells lacking CD20. CD20-expressing T cells respond variably to immunomodulatory treatments given to MS patients: they are reduced by fingolimod, alemtuzumab, and dimethyl fumarate, whereas natalizumab disproportionally increases them in the blood. After depletion by rituximab, they show earlier and higher repopulation than CD20+ B cells. Taken together, human CD3+CD20+ T cells pervade lymphatic organs and the cerebrospinal fluid, have a strong ability to produce different cytokines, and respond to MS disease modifying drugs.

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CD3+CD20+ cells may be an artifact of flow cytometry: Comment on the article by Wilk et al

Cited by 23 publications

References 11 publications

Frequency of Th17 CD20+ cells in the peripheral blood of rheumatoid arthritis patients is higher compared to healthy subjects

Frequency of Th17 CD20+ cells in the peripheral blood of rheumatoid arthritis patients is higher compared to healthy subjects

Rituximab Efficiently Depletes Increased CD20-Expressing T Cells in Multiple Sclerosis Patients

Features of Human CD3+CD20+ T Cells

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