Anti-Human CD40 [Clone HB-14] — Purified in vivo PLATINUM^TM Functional Grade

Clone HB-14 is a monoclonal antibody against human CD40. The most common in vivo applications of clone HB-14 are for studies involving human immune cells (especially B cells and antigen-presenting cells) in humanized mouse models or xenograft systems, rather than in wild-type mice. HB-14 does not recognize mouse CD40, so it is not used for direct targeting of mouse cells in immunocompetent mouse strains.

Key in vivo uses of clone HB-14 in mice include:

• Activation of human B cells and antigen-presenting cells in humanized mice (mice engrafted with human hematopoietic cells, tissues, or PBMCs), aiding research into immune activation, tolerance, and therapies.
• Proliferation assays, where HB-14 can stimulate proliferation of human B cells in vivo or in ex vivo cultures after retrieval from engrafted animals.
• Blockade of CD40-CD40L interaction on human cells, which is important for dissecting mechanisms of immune modulation, transplantation, or autoimmune disease in humanized mice.

Important limitations and context:

• HB-14 is specific for human CD40, so any use in mice requires that the mice have been engrafted with human cells or tissues expressing CD40.
• There is no evidence or published support for using HB-14 directly in standard (non-humanized) mouse models, since mouse CD40 will not be targeted.
• Typical in vivo doses reported for anti-CD40 antibodies range from 200–500 µg per mouse, administered intraperitoneally every 3 days during experimental protocols in humanized mice.

Summary Table:

In standard laboratory mice (not humanized), HB-14 does not have an in vivo application, as its target is not present. If your goal is to manipulate mouse CD40 or mouse immune cells, a different anti-mouse CD40 antibody must be used.

Commonly Used Antibodies and Proteins with HB-14 in Research

The monoclonal antibody HB-14 targets human CD40, a critical immune co-stimulatory molecule expressed on B cells, macrophages, dendritic cells, and other immune cell types. In the literature, HB-14 is frequently used in combination with other antibodies and cytokines to study B cell activation, proliferation, and signaling.

Key Combinations with HB-14

Anti-IgM (Immunoglobulin M):
HB-14 is often paired with antibodies targeting the B cell receptor (BCR), such as anti-IgM, to synergistically enhance B cell proliferation and activation. This combination mimics the physiological scenario where B cells receive signals through both the BCR and CD40, leading to more robust cellular responses than either stimulus alone.

Interleukin-4 (IL-4):
IL-4 is a cytokine commonly used alongside HB-14 to promote B cell differentiation and proliferation. The combination of CD40 ligation (via HB-14) and IL-4 signaling drives B cells toward specific differentiation pathways, including isotype class-switching.

Phorbol 12-myristate 13-acetate (PMA):
PMA, a protein kinase C (PKC) activator, is also used in conjunction with HB-14 to further induce B cell proliferation in experimental settings. This combination is typically employed to test maximal stimulation conditions in vitro.

Functional Effects

• B Cell Proliferation: HB-14, in combination with anti-IgM, IL-4, or PMA, has been shown to significantly promote B cell proliferation in vitro.
• Partial Blockade: While HB-14 can activate CD40 signaling, it also partially blocks the binding of CD40 to its natural ligand, CD40L (CD154), which can be leveraged to dissect signaling pathways.
• T Cell Activation: In some contexts, HB-14 enhances T cell activation, particularly in studies involving HIV-specific CD4+ T cells.

Table of Common Combinations

Summary

In the literature, HB-14 is most commonly used with anti-IgM to enhance B cell receptor signaling, IL-4 to drive differentiation and proliferation, and PMA for maximal activation. These combinations are foundational for studying B cell biology, immune regulation, and CD40-mediated signaling pathways.

Clone HB-14 is primarily associated with the CD40 antibody, which is used in various scientific studies. Here are some key findings related to clone HB-14:

1. CD40 Interaction and Function: Clone HB-14 is a monoclonal antibody that reacts with human CD40, a receptor crucial for B cell differentiation and proliferation, isotype class-switching, and protection against apoptosis. It blocks the interaction between CD40 and its ligand CD154, which is important in T cell-B cell interactions and immune regulation.

2. Applications in Flow Cytometry: Clone HB-14 is commonly used in flow cytometry applications to identify and analyze CD40 expression on cells like B cells, macrophages, and dendritic cells. This antibody is available conjugated with different fluorescent dyes, such as PE and APC, facilitating its use in flow cytometric analyses.

3. Immunological Studies: In immunological research, clone HB-14 can be used to explore the role of CD40 in immune responses, including vaccine studies and investigations into immune regulation mechanisms.

Overall, clone HB-14 is a valuable tool for studying CD40-mediated immune processes and has been utilized in a variety of immunological studies to understand better the role of CD40 in immune response and regulation.

While there are less direct citations specifically mentioning "clone HB-14" in the broader scientific literature outside of its use in CD40 studies, the antibody's role in immunology is well-documented.

Dosing Regimen of Clone HB-14 Across Mouse Models

Summary of Published Evidence

Currently, there is no detailed, peer-reviewed literature in the search results describing the dosing regimens for anti-human CD40 (clone HB-14) antibody specifically across multiple mouse models. The only available information on dosing comes from the manufacturer Leinco, which provides a general dosing guideline but not a model-by-model breakdown.

Leinco’s General Dosing Recommendation

• Dosing Schedule: Every 3–4 days, or 2–3 times per week, for a total of 3–4 doses.
• Purpose: This regimen is intended to ensure adequate receptor saturation and sustained blockade during experiments.

Lack of Model-Specific Dosing Information

The provided sources do not detail how dosing regimens for HB-14 might differ in, for example, syngeneic, xenograft, humanized, or genetically engineered mouse models. Published protocols and reviews for various mouse models (e.g., humanized, orthotopic, chronic infection, hematopoietic stem cell gene therapy) do not mention clone HB-14, nor do they provide comparative dosing tables for this antibody.

Contextual Discussion

The efficacy, pharmacokinetics, and toxicity of HB-14 could theoretically differ between mouse models due to variables such as target expression, immune status, tumor burden, and route of administration. However, without published studies directly comparing HB-14 dosing in different mouse models, any discussion of regimen variation would be speculative.

Clinical Translation and Other Antibodies

Some sources note that, in general, antibody dosing regimens can vary widely between mice and humans, often being more frequent in mice due to faster clearance. This may apply to HB-14, but again, specific data are lacking.

Conclusion

The dosing regimen for clone HB-14 (anti-human CD40) in mice is generally every 3–4 days for 3–4 doses, as recommended by the supplier. There is no published evidence in the search results describing how this regimen varies across different mouse models. Researchers should consult primary literature or directly contact the supplier for model-specific optimization, and may need to perform pharmacokinetic/pharmacodynamic studies tailored to their experimental system.