Bio Tcr Bcr Analysis Scirpy Analysis

Version Compatibility

Reference examples tested with: MiXCR 4.6+, VDJtools 1.2.1+, scanpy 1.10+

Before using code patterns, verify installed versions match. If versions differ:

• Python: pip show <package> then help(module.function) to check signatures

If code throws ImportError, AttributeError, or TypeError, introspect the installed package and adapt the example to match the actual API rather than retrying.

scirpy Analysis

"Analyze single-cell TCR/BCR with gene expression" → Integrate immune receptor clonotype data with scRNA-seq gene expression for joint analysis of clonal expansion and cell state.

• Python: scirpy.io.read_10x_vdj(), scirpy.tl.clonal_expansion(), scirpy.tl.clonotype_network()

Load VDJ Data

Goal: Import single-cell VDJ annotations and integrate them with an existing scRNA-seq AnnData object.

Approach: Read 10x filtered_contig_annotations or AIRR-format files and attach receptor metadata to the AnnData obs.

import scirpy as ir
import scanpy as sc

# Load 10x VDJ data
adata = sc.read_h5ad('scrnaseq.h5ad')

# Add VDJ annotations from 10x filtered_contig_annotations.csv
ir.io.read_10x_vdj(adata, 'filtered_contig_annotations.csv')

# Or load from AIRR format
ir.io.read_airr(adata, 'airr_rearrangement.tsv')

Quality Control

Goal: Identify cells with aberrant chain pairing (doublets, orphan chains, ambiguous pairings).

Approach: Run scirpy chain QC to categorize cells by receptor chain status and visualize QC distributions.

# QC for receptor chains
ir.tl.chain_qc(adata)

# QC categories:
# - multichain: More than 2 chains (potential doublet)
# - orphan: Only one chain detected
# - extra: Extra chains beyond expected pair
# - ambiguous: Ambiguous chain pairing

# Plot QC
ir.pl.group_abundance(adata, groupby='chain_pairing', target_col='receptor_subtype')

Define Clonotypes

# Define clonotypes by CDR3 sequence identity
ir.pp.ir_dist(
    adata,
    metric='identity',
    sequence='aa',
    cutoff=0
)

ir.tl.define_clonotypes(adata, receptor_arms='all', dual_ir='primary_only')

# Check clonotype distribution
print(f"Unique clonotypes: {adata.obs['clone_id'].nunique()}")

Clonal Expansion

# Identify expanded clonotypes
ir.tl.clonal_expansion(adata)

# Categories: 1 (singleton), 2, 3-10, >10

# Plot expansion by cell type
ir.pl.clonal_expansion(adata, groupby='cell_type')

Repertoire Diversity

# Calculate diversity metrics per group
diversity = ir.tl.repertoire_overlap(
    adata,
    groupby='sample',
    target_col='clone_id',
    metric='jaccard'
)

# Alpha diversity
ir.tl.alpha_diversity(adata, groupby='sample', target_col='clone_id')

Compare Groups

# Compare clonotype sharing between groups
ir.pl.group_abundance(
    adata,
    groupby='clone_id',
    target_col='condition',
    max_cols=20
)

# Repertoire overlap heatmap
ir.pl.repertoire_overlap(adata, groupby='sample', target_col='clone_id')

V(D)J Gene Usage

# Plot V gene usage
ir.pl.vdj_usage(
    adata,
    vdj_cols=['v_call_TRA', 'v_call_TRB'],
    full_names=False
)

# Spectratype (CDR3 length distribution)
ir.pl.spectratype(adata, chain='TRB', target_col='cell_type')

Integration with Gene Expression

# Subset to cells with TCR
adata_tcr = adata[adata.obs['has_ir'] == 'True'].copy()

# Find marker genes for expanded vs non-expanded
adata_tcr.obs['is_expanded'] = adata_tcr.obs['clonal_expansion'].isin(['3-10', '>10'])

sc.tl.rank_genes_groups(adata_tcr, groupby='is_expanded')
sc.pl.rank_genes_groups(adata_tcr, n_genes=20)

# UMAP colored by clonal expansion
sc.pl.umap(adata_tcr, color=['cell_type', 'clonal_expansion'])

Export for Downstream Analysis

# Export clonotype table
clonotypes = adata.obs[['clone_id', 'IR_VDJ_1_junction_aa', 'IR_VJ_1_junction_aa',
                         'IR_VDJ_1_v_call', 'IR_VDJ_1_j_call']].drop_duplicates()
clonotypes.to_csv('clonotypes.csv')

# Export for VDJtools
ir.io.write_airr(adata, 'scirpy_airr.tsv')

Related Skills

• mixcr-analysis - Process raw VDJ FASTQ
• single-cell/data-io - Load scRNA-seq data
• single-cell/clustering - Cell type annotation