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Journal for immunotherapy of cancer2021; 9(9); doi: 10.1136/jitc-2021-002688

Simple and effective bacterial-based intratumoral cancer immunotherapy.

Abstract: We describe intratumoral injection of a slow-release emulsion of killed mycobacteria (complete Freund's adjuvant (CFA)) in three preclinical species and in human cancer patients. Efficacy and safety were tested in mammary tumors in mice, in mastocytomas in mice and dogs, and in equine melanomas. In mice, survival, tumor growth, and tumor infiltration by six immune cell subsets (by flow cytometry) were investigated and analyzed using Cox proportional hazards, a random slopes model, and a full factorial model, respectively. Tumor growth and histology were investigated in dogs and horses, as well as survival and tumor immunohistochemistry in dogs. Tumor biopsies were taken from human cancer patients on day 5 (all patients) and day 28 (some patients) of treatment and analyzed by histology. CT scans are provided from one patient. Significantly extended survival was observed in mouse P815 and 4T1 tumor models. Complete tumor regressions were observed in all three non-human species (6/186 (3%) of mouse mastocytomas; 3/14 (21%) of canine mastocytomas and 2/11 (18%) of equine melanomas). Evidence of systemic immune responses (regression of non-injected metastases) was also observed. Analysis of immune cells infiltrating mastocytoma tumors in mice showed that early neutrophil infiltration was predictive of treatment benefit. Analysis of the site of mastocytoma regression in dogs weeks or months after treatment demonstrated increased B and T cell infiltrates. Thus, activation of the innate immune system alone may be sufficient for regression of some injected tumors, followed by activation of the acquired immune system which can mediate regression of non-injected metastases. Finally, we report on the use of CFA in 12 human cancer patients. Treatment was well tolerated. CT scans showing tumor regression in a patient with late-stage renal cancer are provided. Our data demonstrate that intratumoral injection of CFA has major antitumor effects in a proportion of treated animals and is safe for use in human cancer patients. Further trials in human cancer patients are therefore warranted. Our novel treatment provides a simple and inexpensive cancer immunotherapy, immediately applicable to a wide range of solid tumors, and is suitable to patients in developing countries and advanced care settings.
© Author(s) (or their employer(s)) 2021. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ.
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Publication Date: 2021-09-18 PubMed ID: 34531247PubMed Central: PMC8449973DOI: 10.1136/jitc-2021-002688Google Scholar: Lookup
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  • Journal Article
  • Research Support
  • Non-U.S. Gov't

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

This research study details the effectiveness of using killed mycobacteria, in the form of complete Freund’s adjuvant (CFA), for intratumoral injections in various species and human cancer patients. The researchers found that the treatment can lead to tumor regression, survival extension, and is safe for human use.

Study Design and Methodology

  • The researchers conducted their study using three different preclinical animal species and human cancer patients. The animal models used were mice with mammary tumors and mastocytomas, dogs with mastocytomas, and horses with melanomas.
  • Essentially, the killed mycobacteria were prepared in a slow-release emulsion form called complete Freund’s adjuvant (CFA) and injected directly into the tumors.
  • The treatment’s effectiveness was assessed by watching for changes in survival rates, tumor growth, and the infiltration of immune cells in the tumor in mice. In dogs and horses, tumor growth, histology, and survival rates were examined.
  • Human cancer patients underwent tumor biopsies on the 5th and 28th day of treatment, after which the biopsies were analyzed histologically. One patient’s CT scans were also reviewed.

Findings in Animal Models

  • The treatment application extended survival in mice tumor models significantly.
  • There was a complete regression of tumors across all three animal models, albeit with varying success rates. Mouse mastocytomas had 3% regression, canine mastocytomas had 21%, and equine melanomas had 18% regression.
  • The researchers also noted evidence of systemic immune responses by observing the regression of non-injected metastases.
  • Analysis of immune cell infiltration hinted that early infiltration of neutrophils might predict positive treatment outcomes. Further analysis showed increased B and T cell infiltration weeks or months after treatment at the site of canine mastocytoma regression.

Findings in Human Patients

  • The researchers moved to test the adequacy of CFA in 12 human cancer patients, primarily they were looking at the safety of the treatment. The treatment was well tolerated among the patients.
  • CT scans from one patient with late-stage renal cancer showed tumor regression, demonstrating the efficacy of the treatment in humans alongside its safety.

Implications and Future Directions

  • The success of this study thus shows that CFA could potentially play a significant role in intratumoral cancer immunotherapy. However, more trials involving human patients are necessary to solidify this notion.
  • CFA’s success as a simple, cheap, and effective cancer treatment can greatly benefit many patients, particularly in regions with underdeveloped healthcare systems or advanced care settings.

Cite This Article

APA
Carroll CSE, Andrew ER, Malik L, Elliott KF, Brennan M, Meyer J, Hintze A, Almonte AA, Lappin C, MacPherson P, Schulte KM, Dahlstrom JE, Tamhane R, Neeman T, Herbert EW, Orange M, Yip D, Allavena R, Fahrer AM. (2021). Simple and effective bacterial-based intratumoral cancer immunotherapy. J Immunother Cancer, 9(9). https://doi.org/10.1136/jitc-2021-002688

Publication

Journal for immunotherapy of cancer
ISSN: 2051-1426
NlmUniqueID: 101620585
Country: England
Language: English
Volume: 9
Issue: 9

Researcher Affiliations

Carroll, Christina S E
  • Research School of Biology, Australian National University, Canberra, Australian Capital Territory, Australia.
Andrew, Erin R
  • Research School of Biology, Australian National University, Canberra, Australian Capital Territory, Australia.
Malik, Laeeq
  • Department of Medical Oncology, Canberra Hospital, Canberra, Australian Capital Territory, Australia.
  • Medical School, Australian National University, Canberra, Australian Capital Territory, Australia.
Elliott, Kathryn F
  • School of Veterinary Science, The University of Queensland, Gatton, Queensland, Australia.
Brennan, Moira
  • School of Veterinary Science, The University of Queensland, Gatton, Queensland, Australia.
Meyer, James
  • Adelaide Plains Equine Clinic, Gawler, South Australia, Australia.
Hintze, Alexander
  • Klinik Arlesheim, Haus Wegman, Arlseheim, Switzerland.
Almonte, Andrew A
  • Research School of Biology, Australian National University, Canberra, Australian Capital Territory, Australia.
Lappin, Cassandra
  • School of Veterinary Science, The University of Queensland, Gatton, Queensland, Australia.
MacPherson, Philip
  • School of Veterinary Science, The University of Queensland, Gatton, Queensland, Australia.
Schulte, Klaus-Martin
  • Medical School, Australian National University, Canberra, Australian Capital Territory, Australia.
Dahlstrom, Jane E
  • Medical School, Australian National University, Canberra, Australian Capital Territory, Australia.
  • ACT Pathology, Canberra Hospital, Canberra, Australian Capital Territory, Australia.
Tamhane, Rohit
  • Canberra Imaging Group, Canberra, Australian Capital Territory, Australia.
Neeman, Teresa
  • Biological Data Science Institute, Australian National University, Canberra, Australian Capital Territory, Australia.
Herbert, Elizabeth W
  • Adelaide Plains Equine Clinic, Gawler, South Australia, Australia.
Orange, Maurice
  • Klinik Arlesheim, Haus Wegman, Arlseheim, Switzerland.
Yip, Desmond
  • Department of Medical Oncology, Canberra Hospital, Canberra, Australian Capital Territory, Australia.
  • Medical School, Australian National University, Canberra, Australian Capital Territory, Australia.
Allavena, Rachel
  • School of Veterinary Science, The University of Queensland, Gatton, Queensland, Australia.
Fahrer, Aude M
  • Research School of Biology, Australian National University, Canberra, Australian Capital Territory, Australia aude.fahrer@anu.edu.au.

MeSH Terms

  • Animals
  • Cell Line, Tumor
  • Dogs
  • Female
  • Horses
  • Humans
  • Immunotherapy / methods
  • Male
  • Mice
  • Neoplasms / drug therapy

Conflict of Interest Statement

Competing interests: None declared.

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