Important roles of humoral tumor immunity are often pointed out; however, precise profiles of dominant antigens and developmental mechanisms remain elusive. We systematically investigated the humoral antigens of dominant intratumor immunoglobulin clones found in human cancers. We found that approximately half of the corresponding antigens were restricted to strongly and densely negatively charged polymers, resulting in simultaneous reactivities of the antibodies to both densely sulfated glycosaminoglycans (dsGAGs) and nucleic acids (NAs). These anti‐dsGAG/NA antibodies matured and expanded via intratumoral immunological driving force of innate immunity via NAs. These human cancer–derived antibodies exhibited acidic pH–selective affinity across both antigens and showed specific reactivity to diverse spectrums of human tumor cells. The antibody‐drug conjugate exerted therapeutic effects against multiple cancers in vivo by targeting cell surface dsGAG antigens. This study reveals that intratumoral immunological reactions propagate tumor‐oriented immunoglobulin clones and demonstrates a new therapeutic modality for the universal treatment of human malignancies.

Resistance to immune-checkpoint blockade remains challenging in patients with non-small cell lung cancer (NSCLC). Tumor-infiltrating leukocyte (TIL) quantity, composition, and activation status profoundly influence responsiveness to cancer immunotherapy. This study examined the immune landscape in the NSCLC tumor microenvironment by analyzing TIL profiles of 281 fresh resected NSCLC tissues. Unsupervised clustering based on numbers and percentages of 30 TIL types classified adenocarcinoma (LUAD) and squamous cell carcinoma (LUSQ) into the cold-, myeloid cell-, and CD8+ T cell-dominant subtypes. These were significantly correlated with patient prognosis; the myeloid cell subtype had worse outcomes than the others. Integrated genomic and transcriptomic analyses, including RNA sequencing, whole-exome sequencing, T cell receptor repertoire, and metabolomics of tumor tissue, revealed that immune reaction-related signaling pathways were inactivated, while the glycolysis and K-ras signaling pathways activated in LUAD and LUSQ myeloid cell-subtypes. Cases with ALK and ROS1 fusion genes were enriched in the LUAD myeloid subtype, and the frequency of TERT copy number variations was higher in LUSQ myeloid subtype than in the others. These classifications of NSCLC based on TIL status may be useful for developing personalized immune therapies for NSCLC.

<p>Patient outcomes in immune subtypes. <b>A</b> and <b>B,</b> Kaplan–Meier EFS curves for subtypes of LUAD (A) and LUSQ (B). <i>P</i> values were calculated by multivariate Cox regression. <b>C</b> and <b>D,</b> Relationships of cell density (left) with �45 (right) for each immune cell type infiltrated in tumors to patient prognosis in LUAD (C) and LUSQ (D). Z-scores from Cox proportional hazards analysis are plotted; in the plots, red dots indicate <i>P</i> ≤ 0.05. <b>E</b> and <b>F,</b> Differences in histopathologic findings for immune subtypes of LUAD (E) and LUS (F). The <i>P</i> values determined using Fisher exact test w are plotted; in the plots, red circles indicate <i>P</i> ≤ 0.05 and filled red circles indicate Holm-adjusted <i>P</i> ≤ 0.2. <b>G</b> and <b>H,</b> Frequencies of each classification in immune subtypes in LUAD (G) and LUSQ (H). Numbers of patients are indicated in the bar segments.</p>

<p>Activated and suppressed pathways identified by GSEA using the hallmark gene set in immune subtypes. <b>A</b> and <b>B,</b> Heat maps representing top scored pathways enriched in genes with expression increased and decreased in common in LUAD (A) and LUSQ (B). <b>C</b> and <b>D,</b> Top signatures of pathways with increased expression are shown in red; those for genes with decreased expression are shown in blue. Running enrichment scores for glycolysis, IFNα, IFNγ, K-<i>ras</i> signaling, and TGFβ signaling signatures as commonly activated pathways in immune subtypes of LUAD (C) and LUSQ (D).</p>

<p>Detailed methods to establish B16-F10 subclones and IL33 transfectants, and materials for RT-PCR, gene knockdown, and flow cytometry</p>