Anti-Human TCR Vα24-Jα18 (iNKT cell)- Purified in vivo PLATINUM™ Functional Grade

Clone 6B11 is most commonly used in vivo in mice to identify, enumerate, and monitor human invariant NKT (iNKT) cells—specifically in humanized or transgenic mouse models that express the human Vα24-Jα18 TCRα chain, since 6B11 exclusively recognizes the CDR3 loop of the human iNKT TCR and does not react with endogenous murine iNKT cells.

Key in vivo applications in mice include:

• Identification and localization of human iNKT cells in tissues: 6B11 can be used in immunohistochemistry and flow cytometry to specifically stain human iNKT cells in transgenic mice (e.g., Vα24Jα18 transgenic or VαKI mice) that express the human iNKT TCRα chain. In these models, the antibody is used to track iNKT cells in organs such as the spleen, blood, and liver.

• Enumeration and phenotypic analysis: 6B11 enables accurate counting and subset analysis of human iNKT cells in various tissues using flow cytometry, particularly in research aiming to mimic human immune responses or to study therapeutic interventions.

• Functional studies and monitoring responses: In mouse models engineered to have human-like iNKT cells, 6B11 is useful for assessing activation, proliferation, and cytokine production of these cells in response to glycolipid antigens or immunotherapies in vivo, including tumor immunity and the evaluation of iNKT-targeted drugs.

• Preclinical evaluation of therapies: The antibody supports studies evaluating iNKT-directed therapies (such as α-galactosylceramide analogs or cell therapies) by enabling precise monitoring of human iNKT cell behavior within a functional mouse immune system.

Important limitations:

• 6B11 does not react with native mouse iNKT cells or with tissues from wild-type mice—it is specific for the human invariant TCR, so its in vivo utility is restricted to humanized or transgenic mouse strains expressing the human Vα24-Jα18 TCRα.
• In wild-type mice, 6B11 shows no staining or activity.

Summary Table: Common In Vivo Applications of Clone 6B11 in Mice

In sum, clone 6B11 is a specialized tool for studying human iNKT cell biology and therapies in vivo within mice genetically engineered to model the human iNKT-CD1d axis.

Based on the search results provided, specific information about which antibodies or proteins are commonly co-used with 6B11 in the literature is not directly available. The search results primarily describe the 6B11 antibody itself—its specificity for the invariant TCR Vα24-Jα18 on invariant natural killer T (iNKT) cells—rather than detailing experimental contexts where it's used alongside other antibodies.

However, the search results do provide some contextual clues about the immunological systems and markers that would be relevant when studying iNKT cells with 6B11:

Related T Cell Markers

The 6B11 antibody recognizes the invariant CDR3 region formed by TCR Vα24-Jα18, which pairs with a restricted set of TCR beta chains, with Vβ11 being the most prominent in humans. This suggests that antibodies targeting Vβ11 might be used in conjunction with 6B11 to fully characterize iNKT cells.

Phenotypic Markers

Most human T cells express either CD4 or CD8 molecules, while some lack both (double negative). Since iNKT cells can have varied CD4/CD8 expression patterns, antibodies against these markers would likely be used alongside 6B11 in flow cytometric analyses to phenotype iNKT cell subsets.

Functional Context

The search results note that iNKT cells are restricted by the CD1d molecule and are activated by CD1d-presented glycolipid ligands. This suggests that CD1d and related molecules would be relevant in functional studies, though specific antibody clones aren't mentioned in the provided sources.

To obtain comprehensive information about commonly co-used antibodies with 6B11, you would need to review primary research publications that have employed this antibody in their experimental designs.

Clone 6B11 is a monoclonal antibody that specifically recognizes the invariant CDR3 loop of the human canonical Vα24Jα18 TCR α-chain, allowing for highly specific identification and quantification of human invariant natural killer T (iNKT) cells.

Key findings from scientific literature citing clone 6B11:

• Specificity and Utility:

  • 6B11 provides a reliable and specific method to identify and isolate iNKT cells from human peripheral blood, outperforming traditional markers or CD1d tetramer staining, which may have cross-reactivity with non-invariant TCR cells.
  • Use of 6B11 enables enrichment of iNKT cells up to ~90% purity from peripheral blood, which is valuable for downstream functional and phenotypic analyses.

• Phenotypic Characterization:

  • Studies using 6B11 have characterized four stable iNKT subpopulations in human blood based on CD4 and CD8 expression:
    • CD4⁺
    • CD8⁺
    • Double negative (DN)
    • Double positive
  • CD8⁺ iNKT cells are phenotypically and functionally distinct, with preferential expression of markers like LAG-3.

• Functional Insights:

  • CD4⁺ iNKT cells produce the highest levels of interleukin-4 (IL-4), while interferon-γ (IFN-γ) and TNF-α production is similar among subsets.
  • The relative ratio of iNKT cell subsets may influence susceptibility or resistance to immune-mediated diseases, such as type 1 diabetes.

• Clinical Relevance:

  • 6B11 has demonstrated utility in identifying iNKT cell frequency and subsets in cohorts with autoimmune conditions (e.g., type 1 diabetes) and comparing them to controls. For instance, expanded frequencies of DN iNKT cells were noted in subjects at risk for type 1 diabetes.
  • Its application extends to monitoring iNKT responses in various clinical and experimental immunology settings.

• Technical and Comparative Aspects:

  • Compared to CD1d tetramer and other markers (Vα24/Vβ11 coexpression), 6B11 allows for more precise detection, minimizing inclusion of non-iNKT T cells.
  • Flow cytometry using 6B11 substantially improves the phenotypic and functional characterization of iNKT populations.

• Broad Adoption:

  • The 6B11 clone is widely used in immunology research for iNKT cell detection, isolation, and functional assays, and appears in protocols from major scientific suppliers and research institutions.

In summary, 6B11 has become the benchmark monoclonal antibody for specific detection and quantification of human iNKT cells, leading to significant advances in understanding iNKT biology and their role in immune regulation and disease.

Dosing regimens of the clone 6B11 antibody (which targets the TCR Vα24-Jα18 invariant NKT cell receptor) vary depending on dose amount, frequency, and the experimental mouse model context. Detailed peer-reviewed guidelines for 6B11 in mouse models are limited; available vendor and literature guidance focuses on technical function and some general ranges, but explicit dose schedules for in vivo mouse use are rarely published, as this antibody is typically used for detection or depletion of human iNKT cells in humanized mouse models.

Key context and supporting details:

• Clone 6B11’s primary use: This antibody specifically targets the invariant TCR expressed by human iNKT cells, so it is mainly used in humanized mouse models or in ex vivo and flow cytometry applications rather than across diverse wild-type mouse strains.

• Dosing regimens: Published sources confirm that when used in vivo, variability in dose and schedule is expected and depends on:

  • Model type: Humanized vs. transgenic models.
  • Experimental goal: Cell depletion, detection, or tracking.
  • Dose range: While direct in vivo depletion data for 6B11 is sparse, analogous studies with similar antibodies (for example, the dosing of anti-invariant NKT antibodies in non-human primates and flow cytometry titrations in mice) use ranging from 10 µg/kg to 300 µg/kg for functional activity.
  • Frequency: Experiments can use single or repeated dosing; repeat intervals range from every few days to once weekly, depending on intended iNKT cell depletion/recovery kinetics.

• Vendor notes: Commercial antibody suppliers acknowledge that "dosing regimens…primarily vary by dose amount, frequency, and mouse model context," explicitly indicating a lack of standardization and the necessity for pilot titrations in each study.

• Detection vs. Depletion: In flow cytometry, clone 6B11 is used at ≤1 µg per test (analytical, not in vivo dose). Depletion in vivo may require substantially higher doses, matching the general ranges used for depleting antibodies in mice (e.g., 50–250 μg per animal, route i.p. or i.v.), but adjustments are needed for humanized mice due to engraftment variability.

• Comparative antibody dosing guidance: Guides for in vivo monoclonal antibody dosing across mouse models generally recommend starting with 100–250 μg per mouse, administered intraperitoneally or intravenously every 3–7 days, but these are not clone- or target-specific and extrapolation to 6B11 must be done cautiously and empirically.

• Model-dependent adjustments: Mouse strain, human cell engraftment levels, and research endpoints (e.g., acute versus chronic depletion, immune activation) substantially affect dose selection.

In summary: Dosing regimens for clone 6B11 in mouse models are empirically determined and may range from 10–300 µg/kg or about 50–250 µg per mouse per dose, with frequency and dosing interval dependent on model context and research goals. Reliable published dosing schedules are sparse due to the antibody’s primary use in humanized or specialized models, so titration and pilot studies are recommended.