The effect of blockade of the CD40-CD40L pathway on T and B cell mediated alloreactivity after kidney transplantation

The effect of blockade of the CD40-CD40L pathway on T and B cell mediated alloreactivity after kidney transplantation - Effect of blockade of CD40 on alloreactivity after kidney transplantation

Status

Active, not recruiting

Phases

Unknown

Study type

Observational

Source

NL-OMON

Registry ID

NL-OMON42873

Enrollment

Registered

2016-08-05

Start date

2016-08-05

Completion date

Unknown

Last updated

2024-03-11

For informational purposes only — not medical advice. Sourced from public registries and may not reflect the latest updates. Terms

Conditions

orgaantransplantatie kidney transplantation organ transplantation

Interventions

None listed

Eligibility

Age

18 Years to 99 Years

Inclusion criteria

Inclusion criteria: All patients participating in the main trail (MEC 2015-730)

Exclusion criteria

Exclusion criteria: None

Design outcomes

Primary

Measure	Time frame
Peripheral Blood 1. The number and characteristics of peripheral effector and regulatory T-, and B- cells and dendritic cells (DC) of CFZ533-treated patients a. Multicolor flow cytometry to determine the number and characteristics of circulating T and B cells using the One study protocol in combination with intracellular cytokine measurements to determine the frequency of IFN-γ, IL-21 and IL-10 producing T and B cells. 2. To analyze T-B cell interaction during co-stimulation blockade a. Pure cell populations will be sorted with FACS Aria Cell SorterTM: CD4+CXCR5+ T-cells (Tfh cells)and CD19+CD27+ B cells (memory B cells). These memory B cells will be co-cultured with Tfh in the presence of donor antigen for 7 days to study the maximal Tfh-B cell interaction. Differentiation of B cells into plasmablasts (CD19+IgD-CD27+CD38++ cells) will be determined with flow cytometry. IgM and IgG production will be measured with a sandwich ELISA on the supernatant of the co-cultures. Donor-specific antibodies in culture supernatant by Luminex technology. b. Differentiation of patient derived CD43neg B cells into plasma cells will be studied after stimulation with an agonist CD40, anti-IgM and IL-21 (mimicking T cell stimulation). This assay will determine the functional blockade by the test compound. c. Elispot will be used to determine the frequency of donor-antigen specific Tfh-IL-21 producing cells. The frequency of donor specific IgG producing plasma cells will be counted using Elispot. 3. To study the efficacy of CFZ533 on CD40 receptor expression by peripheral blood cells a. CD40 saturation efficacy on peripheral B cells and DC by flow cytometry Biopsy 4) Characterization and quantification of the cellular infiltrate by immunohistochemistry a. CD3, CD4, CD8, CD79a, CD20, CD19, CD38, granzyme B, TIA1, C4d, C3d, CD40, CD40L, PD1, PD1L, BCL6, FoxP3 b. T and B cell mRNA expression profile by Q-PCR	—

Measure

Time frame

Peripheral Blood 1. The number and characteristics of peripheral effector and regulatory T-, and B- cells and dendritic cells (DC) of CFZ533-treated patients a. Multicolor flow cytometry to determine the number and characteristics of circulating T and B cells using the One study protocol in combination with intracellular cytokine measurements to determine the frequency of IFN-γ, IL-21 and IL-10 producing T and B cells. 2. To analyze T-B cell interaction during co-stimulation blockade a. Pure cell populations will be sorted with FACS Aria Cell SorterTM: CD4+CXCR5+ T-cells (Tfh cells)and CD19+CD27+ B cells (memory B cells). These memory B cells will be co-cultured with Tfh in the presence of donor antigen for 7 days to study the maximal Tfh-B cell interaction. Differentiation of B cells into plasmablasts (CD19+IgD-CD27+CD38++ cells) will be determined with flow cytometry. IgM and IgG production will be measured with a sandwich ELISA on the supernatant of the co-cultures. Donor-specific antibodies in culture supernatant by Luminex technology. b. Differentiation of patient derived CD43neg B cells into plasma cells will be studied after stimulation with an agonist CD40, anti-IgM and IL-21 (mimicking T cell stimulation). This assay will determine the functional blockade by the test compound. c. Elispot will be used to determine the frequency of donor-antigen specific Tfh-IL-21 producing cells. The frequency of donor specific IgG producing plasma cells will be counted using Elispot. 3. To study the efficacy of CFZ533 on CD40 receptor expression by peripheral blood cells a. CD40 saturation efficacy on peripheral B cells and DC by flow cytometry Biopsy 4) Characterization and quantification of the cellular infiltrate by immunohistochemistry a. CD3, CD4, CD8, CD79a, CD20, CD19, CD38, granzyme B, TIA1, C4d, C3d, CD40, CD40L, PD1, PD1L, BCL6, FoxP3 b. T and B cell mRNA expression profile by Q-PCR

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Countries

Netherlands

Outcome results