Consequences of inter cellular molecular transfer in vitro and in vivo
Lead Research Organisation:
King's College London
Department Name: Immunology Infection and Inflam Diseases
Abstract
Cell to cell interaction within the immune system takes place within the immunological synapse. As a consequence, surface molecules can be exchanged. We will use an antibody-directed toxin approach to deplete recipient cells that have acquired donor MHC class II molecules (and vice versa) to study their contribution towards the rejection process in a mouse kidney allograft model. An anti donor MHC class II antibody will be used to direct a 'toxin' (gelonin) towards cells that express donor class II MHC molecules. If the donor (but not recipient) cells are resistant to the toxin, then only recipient (Multi drug resistant gene knockout mice which are toxin sensitive) cells that have acquired donor class II MHC molecules will be lysed. This will enable us to determine the relative contribution of this process to the alloresponse. We have synthesized this immunotoxin and demonstrated its effectiveness both in vitro and in vivo. In vitro experiments will also be performed to investigate the ability of the phenomenon to link the direct and indirect antigen presentation pathways. Understanding the consequences of MHC transfer will also have important implications in other fields where antigen presentation is involved, such as tumor immunology, vaccination, infection and auto-immune diseases.
Technical Summary
Cell to cell interaction within the immune system takes place within the immunological synapse. As a consequence, surface molecules can be exchanged. We will use an antibody-directed toxin approach to deplete recipient cells that have acquired donor MHC class II molecules (and vice versa) to study their contribution towards the rejection process in a mouse kidney allograft model. An anti donor MHC class II antibody will be used to direct a 'toxin' (gelonin) towards cells that express donor class II MHC molecules. If the donor (but not recipient) cells are resistant to the toxin, then only recipient (Multi drug resistant gene knockout mice which are toxin sensitive) cells that have acquired donor class II MHC molecules will be lysed. This will enable us to determine the relative contribution of this process to the alloresponse. We have synthesized this immunotoxin and demonstrated its effectiveness both in vitro and in vivo. In vitro experiments will also be performed to investigate the ability of the phenomenon to link the direct and indirect antigen presentation pathways. Understanding the consequences of MHC transfer will also have important implications in other fields where antigen presentation is involved, such as tumor immunology, vaccination, infection and auto-immune diseases.
Planned Impact
In addition to the potential benefits to academia stated in the previous section, the proposed research may have impact in other ways: 1. Although exchange of cell surface molecules had been know for some time. However, despite research efforts in recent years, its functional consequence in a situation relevant to real life, as oppose to simulated in vitro settings, is still unknown. Models to study this had been lacking as it is not possible to stop intercellular molecular transfer in vivo. The experiments proposed here can address some of these questions that many had not been able to answer for a long time. 2. The model we have devised here using immunotoxin and animals that have enhanced sensitivity to the toxin is novel. We have already demonstrated its effectiveness and can be used in other areas of biomedical research to address different questions.
People |
ORCID iD |
Wilson Wong (Principal Investigator) |
Publications
Brown K
(2016)
Immunotoxin Against a Donor MHC Class II Molecule Induces Indefinite Survival of Murine Kidney Allografts.
in American journal of transplantation : official journal of the American Society of Transplantation and the American Society of Transplant Surgeons
Nowocin AK
(2019)
Characterizing the B-Cell and Humoral Response in Tertiary Lymphoid Organs in Kidney Allografts.
in Experimental and clinical transplantation : official journal of the Middle East Society for Organ Transplantation
Nowocin AK
(2015)
An Extraperitoneal Technique for Murine Heterotopic Cardiac Transplantation.
in American journal of transplantation : official journal of the American Society of Transplantation and the American Society of Transplant Surgeons
Brown K
(2020)
A novel in vivo model using immunotoxin in the absence of p-glycoprotein to achieve ultra selective depletion of target cells: Applications in trogocytosis and beyond.
in Journal of immunological methods
Description | We have developed an immunotoxin which consists of an antibody part that directs a plant toxin towards specific target cells. This was used in an organ transplantation model The immunotoxin directed the toxin to donor cells that are capable of eliciting an immune response. When given on the day of transplantation, the immunotoxin was able to prolong the survival of donor kidney grafts in recipients that would otherwise reject the grafts. There was diminished donor-specific antibody formation and delayed rejection of subsequent donor type skin grafts. This strategy can be adopted to be used in the clinic to prolong survival of donor grafts in humans. In the second part of the project, we used the same immunotoxin to study a specific immunological phenomenon, that of exchange of surface molecules between cells. We have used recipients that are sensitive to killing by the toxin part of the immunotoxin, but they do not express the target molecule that the antibody part of the immunotoxin recognises. Therefore, only recipient cells that have acquired the relevant molecule would be killed. We have successfully demonstrated that this model allows us to deplete this type of cells, and studied the consequences of this. After depletion of recipient cells that have acquired the molecule recognised by the immunotoxin, there was a slight reduction in antibodies produced, but survival of the transplant organs were unaffected. This is a useful tool for highly specific cell depletion and can be used by others in other areas of investigation. |
Exploitation Route | The immunotoxin can be used to prolong survival of donor organs. The method that we have developed to deplete cells are highly specific, this can be used as a fine tool to dissect immunological mechanisms. |
Sectors | Healthcare |
Description | The first key finding has now been published: Brown K, Nowocin AK, Meader L, Edwards LA, Smith RA Wong W. Immunotoxin against a donor MHC class II molecule induces indefinite survival of murine kidney allografts. American Journal of Transplantation. 2016 In press. Early view available at: http://www.ncbi.nlm.nih.gov/pubmed/26799449 We have paid for open access. Therefore, this article is available to anyone free of charge. Dr Wong has been invited to give a talk on this finding at the "The 2016 Immunosuppression Summit: Problems of immunosuppression and overcoming them." A second manuscript detailing the second part of or key finding is being prepared. |
First Year Of Impact | 2016 |
Sector | Pharmaceuticals and Medical Biotechnology |
Impact Types | Economic |
Description | Prolonging cardiac allograft survival by targeting the indirect antigen presentation pathway with an immunotoxin |
Amount | £248,748 (GBP) |
Funding ID | PG/17/52/33059 |
Organisation | British Heart Foundation (BHF) |
Sector | Charity/Non Profit |
Country | United Kingdom |
Start | 02/2018 |
End | 01/2020 |
Title | Novel site for heterotopic mouse heart transplantation |
Description | We have developed a new method for transplanting heart grafts in mice by placing it in the groin instead of the abdomen. This reduces the trauma of the operation, resulting in faster recovery time. |
Type Of Material | Model of mechanisms or symptoms - mammalian in vivo |
Provided To Others? | No |
Impact | This is a refinement, under the 3Rs |
Description | Proteinomics studies in lymphatic fluid following transplantation |
Organisation | King’s Health Partners |
Department | King's Health Partners Academic Health Science Centre (NIHR BRC) |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | We have developed a way to collect lymph fluid following islet transplantation in mice for proteinomics studies. |
Collaborator Contribution | Collaboration with Dr Aileen King at King's College London has enabled us to perform allogeneic Islet transplantation for lymph collection to study the lymph draining from kidneys that has received the donor islets. The proteinomics laboratory will be analysing the samples that we have collected. |
Impact | A full project grant application has been submitted to the BBSRC. |
Start Year | 2014 |
Description | Synthesis of immunotoxin |
Organisation | King's College London |
Department | MRC Centre for Transplantation |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | We have generated an antibody against a donor MHC class II molecule. This will be used to synthesise an immunotoxin to be tested in our in vivo model |
Collaborator Contribution | We are collaborating with Dr Richard Smith. He will be advising us on the synthesis of the immunotoxin. |
Impact | We have established a protocol for the synthesis of the immunotoxin. |
Start Year | 2018 |
Description | Animal research and organ transplants |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | Yes |
Type Of Presentation | Keynote/Invited Speaker |
Geographic Reach | International |
Primary Audience | Public/other audiences |
Results and Impact | It helps the general public to understand why animal research is necessary in transplantation research. The video has been viewed 367 times to date. |
Year(s) Of Engagement Activity | 2013 |
URL | http://www.understandinganimalresearch.org.uk/resources/video-library/46/organ-transplants-and-anima... |
Description | Oral presentation in Congress. Immunotoxin in the Absence of P-Glycoprotein allows Selective Target Cell Killing in the Semi-Direct Antigen Presentation Pathway Following Organ Transplantation |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Professional Practitioners |
Results and Impact | Combining the use of an immunotoxin with MDR-/- hosts allows ultra selective depletion of recipient antigen presenting cells in the semi-direct pathway. This technique may help to elucidate the involvement of this pathway in the rejection response. |
Year(s) Of Engagement Activity | 2018 |
Description | The 2016 Immunosuppression Summit: Problems of immunosuppression and overcoming them |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Professional Practitioners |
Results and Impact | Title of the talk was "Targeting donor rather than recipient cells to prevent rejection of transplanted organs." The event will take place between 15th and 17th March 2016. The full impact has not been recognised yet. |
Year(s) Of Engagement Activity | 2016 |
URL | http://www.Immunosuppression2016.com |