Establishment of anti-mesothelioma monoclonal antibodies

Our plain-English summary of Establishment of anti-mesothelioma monoclonal antibodies, by N. Mizutani, M. Abe, S. Matsuoka, et al (BMC Research Notes, 2016).

The study

• Authors: N. Mizutani, M. Abe, S. Matsuoka, K. Kajino, M. Wakiya, N. Ohtsuji, R. Hatano, C. Morimoto, O. Hino
• Journal: BMC Research Notes
• Published: 2016;9:324
• Cited by: 2 articles, as counted by NIH iCite
• Read it: PubMed · Publisher · Free full text

Abstract

Background: Mesotheliomas are aggressive, therapy-resistant tumors that are predicted to increase in incidence at least until 2020. The prognosis of patients with mesothelioma is generally poor because they are typically diagnosed at a late stage and their tumors are resistant to current conventional therapies. For these reasons, improved diagnosis and therapy are urgently required. To address these issues, the aim of our research was to develop novel mesothelioma-specific monoclonal antibodies (mAbs) as diagnostic and therapeutic agents.

Methods: To develop anti-mesothelioma mAbs useful for diagnosis and therapy, we repeatedly immunized a BALB/c mouse with viable mesothelioma cells, alternating between those from three mesothelioma cell lines. We hybridized the spleen cells from this immunized mouse with P3U1 myeloma cells. We then screened supernatants harvested from the hybridoma clones by assessing whether they bound to a mesothelioma cell line not used for immunization and altered its morphology. We designed this developmental strategy to reduce the risk of obtaining clonotypic mAbs against a single mesothelioma cell line.

Results: Our newly generated mouse anti-human mAbs immunostained clinical samples of mesotheliomas. One of the newly generated mAbs did not react with any other tumor cell line tested. Two other mAbs significantly inhibited the proliferation of mesothelioma cells.

Conclusion: These newly generated anti-mesothelioma mAbs are potentially useful as diagnostic and therapeutic agents for mesothelioma. Moreover, our novel strategy for establishing antitumor mAbs may facilitate the development of new diagnostic and therapeutic techniques for mesotheliomas and other malignancies.

In plain English

This laboratory study aimed to make new antibodies that recognize mesothelioma cells. It did not study asbestos exposure. The authors repeatedly injected one mouse with live cells from three human mesothelioma cell lines. They then picked four antibody-producing clones, JMAM1 to JMAM4, that reacted with a fourth mesothelioma cell line. In lab tests, JMAM1 to JMAM3 bound both epithelial and sarcomatous mesothelioma cell lines; JMAM4 bound only epithelial ones. JMAM4 did not detectably bind any lung or other cancer cell line tested. All four stained cell membranes in pleural fluid samples from three mesothelioma patients. They also stained non-cancerous reactive mesothelial cells in pleural fluid, but not normal pleura or lung tissue, nor formalin-fixed, paraffin-embedded mesothelioma tissue. At the highest dose, JMAM1 and JMAM3 reduced one mesothelioma cell line's growth by at least 50 and 40 percent respectively. The effects of JMAM2 and JMAM4 on growth were not statistically significant. Only JMAM1 significantly reduced cell invasion. The authors concluded that the antibodies are potentially useful for diagnosis and therapy. They noted that the antibodies' targets were not identified.

We wrote this plain-English summary from the full article with the help of AI, and a second AI model checked each sentence against the article. It is not the authors' wording; the abstract and the article are their own account.

What the study did and found

• Design: Laboratory study: one mouse immunized with live human mesothelioma cell lines to produce monoclonal antibodies, which were then tested on cell lines and on pleural effusion samples from patients
• Who was studied: One female BALB/c mouse for immunization; human mesothelioma cell lines and cell lines from lung, breast, liver, pancreatic, gastric, colon, ovarian and renal cancers; pleural effusion samples from three patients with cytologically diagnosed mesothelioma at Juntendo University, Japan
• Exposure: No asbestos exposure was studied. The mouse was immunized with live cells from the mesothelioma cell lines ACC-MESO4, MSTO-211H and NCI-H226, and four resulting antibodies (JMAM1 to JMAM4) were tested
• Main results:
  • In flow cytometry, JMAM1, JMAM2 and JMAM3 stained both epithelial and sarcomatous mesothelioma cell lines. JMAM4 stained the epithelial cell lines only.
  • JMAM4 did not detectably react with any of the non-lung carcinoma cell lines tested, nor with any of the lung cancer cell lines.
  • JMAM1 did not detectably bind the epithelial-type lung cancer cell lines A549 and PC9. It did bind the small-cell lung cancer lines WA-hT and Lu24, and among the other carcinoma lines it reacted only with the renal cell carcinoma line VMRC-RCW.
  • Anti-calretinin antibody partially inhibited JMAM2 staining of NCI-H226 mesothelioma cells.
  • JMAM1 to JMAM4 stained cell membranes in the ethanol-fixed pleural effusion specimens of all three mesothelioma patients.
  • Reactive mesothelial cells in pleural effusion stained with the JMAM antibodies. Normal pleural mesothelium and normal lung tissue did not stain.
  • In MSTO-211H mesothelioma cells, proliferation was reduced by at least 50 percent with 0.4 µg/ml of JMAM1 and by at least 40 percent with 0.4 µg/ml of JMAM3.
  • The proliferation effects of JMAM2 (P = 0.1) and JMAM4 (P = 0.18) were not statistically significant. For JMAM1 and JMAM3, P was less than 0.05.
  • In a wound-healing assay, JMAM1, JMAM2 and JMAM3 significantly inhibited the migration of NCI-H226 mesothelioma cells.
  • In an invasion assay with NCI-H226 cells, only JMAM1 was statistically significant (P less than 0.05). The results for the others were JMAM2 P = 0.12, JMAM3 P = 0.08 and JMAM4 P = 0.18.
• The authors' conclusion: The authors concluded that JMAM1 to JMAM4 are potentially useful as diagnostic and therapeutic agents for mesothelioma, and that their method of making antibodies may help with other cancers as well.
• Limitations the authors state:
  • The authors did not identify the target molecules of the JMAM1 to JMAM4 antibodies.
  • The antibodies did not stain formalin-fixed, paraffin-embedded specimens of surgically resected mesothelioma tissue.

Topics

The National Library of Medicine indexes this study under subjects our site covers: Mesothelioma.

Medical terms in this study, explained in our Lexicon: Mesothelioma, Monoclonal Antibody, Prognosis, Myeloma.

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