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Multi-Omic Integration: TCR × ELISpot

TCR Repertoire Analysis — Notebook 07: Functional immune response correlation, multi-omic composite scoring, and translational implications
Joshua LuthyR + edgeR + tidyverseSynthetic Data2025
Contents
  1. Analysis Overview
  2. ELISpot Response Overview
  3. TCR × ELISpot Integration
  4. Multi-Omic Composite Score
  5. Conclusions

01 Analysis Overview

This notebook integrates TCR repertoire data with ELISpot functional immune response data to build a multi-omic picture of clonal expansion and T cell effector function. By correlating ELISpot responses (IFN-γ, IL-2, Granzyme B) against tumor-associated antigens (WT1, PRAME, Survivin, NY-ESO-1, MAGE-A3) with TCR clonal expansion metrics, we assess whether expanding clonotypes drive measurable anti-tumor immunity.

3
Cytokines Measured
5
Target Antigens
14/30
Positive Responses
5
Timepoints

02 ELISpot Response Overview

Figure 1. Peak IFN-γ ELISpot response (spots/10⁶ cells) by patient × antigen. CR patients show strong multi-antigen responses while PD patients show minimal reactivity.
Finding

CR patients mount positive ELISpot responses against 3–5 antigens while PD patients respond to 0–2. Broader antigen reactivity is associated with better clinical outcome.

03 TCR × ELISpot Integration

We correlate TCR clonal expansion (AUC) with functional ELISpot response (sum IFN-γ) to test whether the clonotypes expanding post-infusion are driving measurable anti-tumor immunity.

Figure 2. Clonal expansion (mean AUC) vs functional immune response (total IFN-γ). Strong positive correlation confirms that expanding clonotypes drive measurable anti-tumor T cell immunity.

04 Multi-Omic Composite Score

We build a final composite score combining both TCR repertoire (clonality, AUC, persistence) and ELISpot (total IFN-γ, antigen breadth) features, weighting each modality equally.

Figure 3. Multi-omic composite score showing TCR (cyan) and ELISpot (green) components. Both modalities contribute to separating CR from PD patients.
Figure 4. Integrated TCR × ELISpot heatmap (z-scored). Both structural (TCR) and functional (ELISpot) metrics converge — CR patients show consistently favorable profiles.

05 Conclusions

Multi-Omic Integration Findings

1. TCR clonal expansion and ELISpot functional responses are strongly correlated — expanding clonotypes drive measurable anti-tumor immunity.

2. Antigen breadth associates with outcome: CR patients react to 3–5 antigens, PD to 0–2.

3. Clonal persistence predicts sustained functional immunity at Wk12.

4. The combined TCR + ELISpot composite score provides better separation of response groups than either modality alone.

5. Both structural (TCR) and functional (ELISpot) arms converge: effective cell therapy is characterized by focused clonal expansion of persistent, functionally active T cells against multiple tumor antigens.

Translational Implications