A Simple and Rapid Protocol to Non-enzymatically Dissociate Fresh Human Tissues for the Analysis of Infiltrating Lymphocytes

Garaud, Soizic; Gu-Trantien, Chunyan; Lodewyckx, Jean Nicolas; Boisson, Anaïs; Silva, Pushpamali De; Buisseret, Laurence; Migliori, Edoardo; Libin, Myriam; Naveaux, Céline; Duvillier, Hugues; Willard-Gallo, Karen

doi:10.3791/52392

Cited by 24 publications

(23 citation statements)

References 15 publications

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“…B cells infiltrating fresh tissue specimens from normal (n = 62), nonadjacent nontumor (NANT; n = 312), benign tumor (n = 21), untreated IDC (n = 241), and untreated invasive lobular carcinoma (ILC; n = 62) were analyzed by FACS ( Figure 2). The clinicopathological characteristics of the patients are detailed in Table 1, and the FACS gating strategies shown are in Supplemental Figure 1 (47). TIL-B were detected at significantly higher densities (absolute number/ mg tissue) and percentages (percentage of total CD45 + cells) in invasive carcinomas (IDC and ILC) compared with normal breast tissues ( Figure 2, A and B).…”

Section: Resultsmentioning

confidence: 99%

“…The functionality of TIL-B in BC. We evaluated the functional attributes of TIL-B, beginning with the balance of Ig isotypes present in fresh breast tissue supernatants (47) and plasma. Plasma from HD and BC patients, the latter stratified on TIL levels, were remarkably similar except for an increase in IgG3 detected in the TIL neg group (Figure 5A).…”

Section: Table 1 Til-b Density Association With Clinicopathological mentioning

confidence: 99%

See 1 more Smart Citation

Tumor-infiltrating B cells signal functional humoral immune responses in breast cancer

Garaud¹,

Buisseret²,

Solinas³

et al. 2019

JCI Insight

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248

165

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Section: Resultsmentioning

confidence: 99%

Section: Table 1 Til-b Density Association With Clinicopathological mentioning

confidence: 99%

Tumor-infiltrating B cells signal functional humoral immune responses in breast cancer

Garaud¹,

Buisseret²,

Solinas³

et al. 2019

JCI Insight

Self Cite

248

165

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“…Fresh breast tumor tissues were collected immediately following surgery and dissociated (without enzymatic digestion) using the gentleMACS Dissociator (Miltenyi Biotec) prior to Ab labeling, as previously described (52 (26). DNA methylation was analyzed on Infinium HumanMethylation450K bead arrays as previously described for Infinium HumanMethylation27K bead arrays (26).…”

Section: Discussionmentioning

confidence: 99%

DNA methylation–based immune response signature improves patient diagnosis in multiple cancers

Jeschke

Bizet

Desmedt

et al. 2017

Journal of Clinical Investigation

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118

102

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“…Matching para-tumor tissues were defined as those ≤ 2.0 cm from the tumor edge and the adjacent normal tissues were procured at the most distant site from the resected specimen (≥ 5.0 cm from the tumor edge). Fresh tissue samples were cut and nonenzymatically dissociated, and single cells were prepared according to a previously described protocol [18]. PBMC were isolated using Ficoll-Paque PLUS (GE Healthcare, USA) according to the manufacturer's instructions.…”

Section: Patients and Sample Preparationmentioning

confidence: 99%

B7H4 expression in tumor cells impairs CD8 T cell responses and tumor immunity

Zhou

Ruan

Liu

et al. 2019

Cancer Immunol Immunother

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B7 homolog 4 (B7H4) is considered a negative regulator of immune responses, but the immunoregulatory role of B7H4 in the tumor microenvironment is not clear. Here, we assessed B7H4 expression cell types in human breast cancer tissues and addressed its potential mechanisms in the CD8 T cell immune response. The results from flow cytometry and immunohistochemistry demonstrated that B7H4 was highly expressed in 26 out of 30 (86.7%) breast invasive ductal carcinomas, and B7H4 surface expression on tumor cells was inversely correlated with CD8 T lymphocytes infiltration (p < 0.0001). In vivo, B7H4-overexpressing tumor cells showed enhanced tumor growth in immunocompetent mice with impaired CD8 T cell infiltration of the tumor. Further investigation showed that activation and expansion of CD8 T cells within the lymph nodes were suppressed in B7H4-overexpessing tumor-bearing mice. An in vitro killing assay showed that the cytotoxicity of CD8 T cells was inhibited in B7H4-overexpressing tumor cells. These findings suggest that B7H4 in tumor cells is a negative regulator of CD8 T cell activation, expansion and cytotoxicity, indicating that tumor cell-associated B7H4 might be a target for T cell-based cancer immunotherapy. Keywords B7 homolog 4 • Tumor microenvironment • T cell • Immune suppression • Adoptive transfer Abbreviations B7H4 B7 homolog 4 CFSE Carboxyfluorescein diacetate succinimidyl ester CTL Cytotoxic T lymphocyte FCM Flow cytometry IHC Immunohistochemistry IDC Invasive ductal carcinomas PBMC Peripheral blood mononuclear cell TCR T cell receptor TAM Tumor associated macrophage TIL Tumor infiltrating lymphocyte * Qiuyu Zhang

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A Simple and Rapid Protocol to Non-enzymatically Dissociate Fresh Human Tissues for the Analysis of Infiltrating Lymphocytes

Cited by 24 publications

References 15 publications

Tumor-infiltrating B cells signal functional humoral immune responses in breast cancer

Tumor-infiltrating B cells signal functional humoral immune responses in breast cancer

DNA methylation–based immune response signature improves patient diagnosis in multiple cancers

B7H4 expression in tumor cells impairs CD8 T cell responses and tumor immunity

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