Clone 10F.9G2 is a rat monoclonal antibody targeting mouse PDL1 (programmed death ligand 1) that has become a widely utilized tool in mouse immunology research. This antibody serves multiple critical functions in in vivo studies, particularly in understanding and manipulating immune checkpoint pathways.

Blocking PDL1 Interactions

The primary in vivo application involves disrupting PDL1 binding to its receptors, specifically PD1 and B71 (CD80). By blocking these interactions, 10F.9G2 effectively removes inhibitory signals that normally suppress T cell activation and proliferation. This blocking mechanism operates in a dosedependent manner and has been validated across numerous experimental systems.

Cancer Immunotherapy Models

In oncology research, 10F.9G2 demonstrates significant antitumor activity. The antibody has been used to transiently arrest tumor growth in mouse models of melanoma by blocking the PDL1/PD1 interaction. This application makes it valuable for studying immune checkpoint blockade strategies and evaluating combination immunotherapy approaches in preclinical cancer models.

Autoimmune Disease Studies

10F.9G2 has proven particularly useful in autoimmune disease research, especially diabetes models. In NOD (nonobese diabetic) mice, administration of this antibody rapidly induces diabetes by unleashing diabetogenic effector T cells. In adoptive transfer experiments using CD8+ T cells from diabetic 8.3 NOD mice, treatment with 10F.9G2 resulted in 100% of recipient mice developing rapidonset diabetes, compared to low incidence in controltreated animals. By day 11 posttransfer, approximately 40% of islets showed insulitis in 10F.9G2treated mice versus less than 4% in controls.

Immune Response Modulation

Beyond specific disease models, 10F.9G2 serves as a general tool for detecting, modulating, or enhancing immune responses in mice. The antibody enables researchers to investigate how PDL1mediated signaling regulates T cell function, inflammation, and tolerance in various physiological and pathological contexts. This includes applications in studying dendritic cell function, T cell activation, and the balance between immune activation and suppression.

Technical Considerations

For in vivo applications, 10F.9G2 is available in specialized formulations designed for animal studies, with high purity (>95%) and low endotoxin levels (less than 1 EU/mg). These quality specifications are essential for minimizing nonspecific inflammatory responses that could confound experimental results.

The most commonly used antibodies or proteins with 10F.9G2 (antimouse PDL1/B7H1) in the literature are those that target related immune checkpoint pathways or immune cell markers, especially antiPD1 antibodies, other PDL1 clones, and cell lineage markers.

Commonly coused antibodies and proteins include:

• AntiPD1 antibodies:

  • Clone 29F.1A12: A widely used antimouse PD1 antibody for in vivo and in vitro blockade.
  • Clone RMP114: Another frequently employed antimouse PD1 antibody.

• Other antiPDL1 antibodies:

  • Clone MIH6: Nearly equivalent to 10F.9G2, often used for comparison or alternate epitope targeting.
  • Clone 10F.5C5: Sometimes used in studies analyzing PDL1 expression in presence of 10F.9G2 or MIH6, as it recognizes a distinct epitope.
  • Clone 2H11: Frequently comentioned with 10F.9G2 in literature; it is used due to its distinct functional properties.

• Immune cell markers:

  • CD3, CD4, CD8: To identify T cells and their subsets in flow cytometry or immunofluorescence panels.
  • CD80 (B71): Targeted in studies examining the blocking of PDL1/B71 interaction by 10F.9G2.
  • CD274: The gene encoding PDL1, relevant for broader antibody panels.

• Other functional and lineage markers:

  • Antibodies against B cells (e.g., CD19), NK cells (e.g., NK1.1), dendritic cells (e.g., CD11c), used for immune profiling alongside PDL1 detection.

Researchers often use combinations of these antibodies to investigate immune checkpoint blockade, cell subset phenotyping, and detailed immune landscape characterization in murine models, particularly in studies of cancer immunotherapy and immune regulation.

Clone 10F.9G2 is a widely used rat monoclonal antibody that specifically targets mouse PDL1 (also known as CD274 or B7H1), and it has been instrumental in elucidating the roles of PDL1 in immune regulation, especially in mouse models. Key findings from scientific literature using citations of 10F.9G2 cluster around the following themes:

• Dual blockade of PDL1 interactions: 10F.9G2 is notable as a “dualblocker” that inhibits both PDL1:PD1 and PDL1:B71 (CD80) interactions. This distinguishes it from some other antiPDL1 antibodies, such as 10F.2H11, which only block PDL1:B71 and not PDL1:PD1. Blocking these pathways is key to disrupting immune checkpoint suppression, thereby enhancing Tcell mediated antitumor immunity.

• Distinct epitope recognition: 10F.9G2 binds a unique epitope on PDL1 that is different from other antiPDL1 clones such as 10F.2H11; this has been demonstrated by competitive binding studies. Additionally, it shares overlapping epitopes with another dualblocker, clone MIH5, but not with 10F.2H11, supporting its dualfunctional blocking profile.

• Functional effects in vivo: In mouse experiments, administration of 10F.9G2 increases infiltration of CD8+ effector T cells into pancreatic islets, exacerbating insulitis in diabetogenic models. This highlights the antibody's impact on Tcell infiltration and autoimmunity when PDL1 pathways are blocked.

• Research and immunotherapy utility: 10F.9G2 is extensively used in:

  • In vivo tumor models to demonstrate the role of PDL1 in tumor immune evasion; blockade by 10F.9G2 transiently arrests tumor growth and enhances antitumor immune responses.
  • Immunophenotyping and mechanistic studies to map the impact of PDL1 pathways on diverse immune cell populations within tumor and nontumor environments.
  • Assays such as flow cytometry, immunofluorescence, and inhibition/blocking studies.

• Biochemical properties: The antibody has high specificity for mouse PDL1, suitable for a broad range of applications, with low endotoxin levels in purified formats suitable for in vivo experiments.

• Product impact: 10F.9G2 is one of the most commonly cited antiPDL1 antibodies for murine research, with over 600 academic citations, reflecting its central role in checkpoint blockade research.

• Clinical translation and drug development: Findings using mouse model blockade with 10F.9G2 have informed the design and expected outcomes of human PDL1 checkpoint inhibitors in cancer immunotherapy.

In summary, clone 10F.9G2 is a benchmark reagent in mouse immunology for probing and blocking PDL1 function, and its dual blockade of PD1 and B71 pathways has provided key insights into immune regulation and the basis for checkpoint inhibitor therapy in cancer.

The dosing regimens for clone 10F.9G2, an antiPDL1 antibody, can vary across different mouse models. However, the standard dose range for this clone is typically between 100250 μg per mouse. This is administered intraperitoneally, and the optimal dosing schedule is usually 23 times per week. The dosing regimens are influenced by the specific application, such as cancer immunotherapy studies or infection models, where the goal is to enhance Tcell responses by blocking the PD1/PDL1 interaction.

One study mentions using a specific dose of 200 μg/mouse for PDL1 blockade using 10F.9G2. Variations in dosing can depend on factors like the model's sensitivity to PDL1 blockade, the presence of other immune modulators, and the desired immune response outcomes.

In general, while the core dosing parameters remain consistent across models, specific dosing frequencies and amounts may be adjusted based on experimental objectives and the immunophenotypic characteristics of the models used, such as MC38 and LLC1 tumor models.