| 1. |
- Zhang, Lu, et al.
(författare)
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Detection of micro-metastases by flow cytometry in lymph nodes from patients with penile cancer
- 2018
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Ingår i: BMC Urology. - : BioMed Central (BMC). - 1471-2490. ; 18:1
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Tidskriftsartikel (refereegranskat)abstract
- BACKGROUND: The tumor draining lymph node concept was first described in penile cancer for staging. Immunohistochemistry and histopathology evaluations are routinely used in clinical practice to examine lymph nodes for metastasis. However, these methods are time-consuming with low diagnostic accuracy and micro-metastases might be missed. In this study, we aim to evaluate detection of metastatic cells in draining lymph nodes by flow cytometry.METHODS: To assess the sensitivity of micro-metastasis detection by FACS (Fluorescence-activated cell sorting), HeLa cells were titrated into Peripheral blood mononuclear cells (PBMCs) and expression of pan-cytokeratin AE1/AE3 was analyzed. Single cell suspensions were separately prepared from 10 regional lymph nodes obtained from 5 patients with invasive penile cancer undergoing radical surgery and lymph node dissection. Lymph node dereived cells were examined for cell surface expression of EpCAM, E-cadherin and intracellular expression of pan-cytokeratin AE1/AE3 by FACS.RESULTS: Ten lymph nodes from 5 penile cancer patients were investigated in a head-to-head comparison between FACS and pathology examination of sections. All metastatic lymph nodes verified by pathology examination were also identified by FACS. Two additional lymph nodes with micro-metastases were diagnosed by FACS only.CONCLUSIONS: FACS analyses of pan-cytokeratin AE1/AE3 stained single cells from tumor draining lymph nodes can be used to detect micro-metastases in patients with penile cancer patients.
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| 2. |
- Zhang, Lu, et al.
(författare)
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Immune responses against autologous tumor and human papilloma virus in lymph nodes from patients with penile cancer
- 2021
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Ingår i: Investigative and Clinical Urology. - : The Korean Urological Association. - 2466-0493 .- 2466-054X. ; 62
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Tidskriftsartikel (refereegranskat)abstract
- PURPOSE: Nearly half of penile cancers are related to human papillomavirus (HPV) infection. Investigations of tumor- and HPV-specific T cell reactivity in regional lymph nodes (LNs) from patients with penile cancer are warranted.MATERIALS AND METHODS: In this study, single-cell suspensions from LNs and peripheral blood from 11 patients with penile cancer were stained with antibodies for lymphocyte markers and analyzed by fluorescence-activated cell sorting (FACS). DNA was extracted from the tumor tissue and HPV status was investigated by PCR.RESULTS: T-cell reactivity against autologous tumor-extract and against the HPV-vaccine Gardasil® was tested by flow-cytometric assay of specific cell-mediated immune response in activated whole blood (FASCIA). CD4+/CD8+ ratios were significantly lower in HPV positive LNs (p<0.05). Immune responses to tumor extract assessed by blast transformation and expansion in vitro, of either CD4+ or CD8+ T-cells, were found in 9 of 13 LNs (69%). 5 of 6 tested patients demonstrated T cell recognition of tumor-associated antigen(s). In HPV-positive patients, dose-dependent T cell responses against L1 (late) HPV proteins (Gardasil vaccine) were demonstrated.CONCLUSIONS: LN-derived T cells from patients with penile cancer recognize tumor antigen(s) and in HPV-positive cases, there is a response against L1 (late) HPV proteins, being constituents of the Gardasil vaccine.
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| 3. |
- Zhang, Lu, et al.
(författare)
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Immune responses against Human Papilloma virus in draining lymph nodes from patients with penile cancer
- 2017
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Ingår i: Scandinavian Journal of Immunology. - : John Wiley & Sons. - 0300-9475 .- 1365-3083. ; 86:4, s. 339-339
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Tidskriftsartikel (övrigt vetenskapligt/konstnärligt)abstract
- Human papillomavirus (HPV) is a DNA virus which accounts for 5.2% of all cancers worldwide and is a well‐known cause of cervical, vulvar, head and neck cancer, but studies on penile cancer (PC) are rare. There are more than 170 HPV strains, where HPV16 is the main strain accounting for 94.7% of all PCs. Additional studies have detected HPV DNA in both primary tumors and metastases, indicating the potential role of HPV in tumor progression. We have investigated immune responses against autologous tumor‐extract in draining lymph nodes (LNs) in colon and urinary bladder cancer. There are several indications that HPV associated malignancies eliciting a virus‐antigen‐specific immune response. HPV E6 and E7 as oncogenes are widely studied in HPV associated cancer. A study comparing different HPV antigen‐specific T cell efficiency in killing autologous HPV positive tumor cells in cervical cancer patients, suggests that L1‐specific CD8+ cytotoxic T lymphocytes (CTLs) are equally effective as E7‐specific CD8+ CTLs. We set out to evaluate immune responses in LNs from PC patients against the HPV vaccine Gardasil (L1). In this study, nearly half of the PC patients were detected as HPV positive by PCR. Phenotypes of the lymphocytes from LNs were characterized. We used FASCIA to investigate lymphocyte responses towards tumor‐extract and Gardasil in LNs, found a good response towards Gardasil among HPV‐positive patients but not in HPV‐negative patients. A dose dependent reactivity was observed. Strategies using HPV L1 proteins as antigens may be useful for adoptive T cell therapy of patients with PC.
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