Short peptides, which often originate from protein processing in tissue,may act as immunomodulators by changing the number and activity of immune cells and thus affect (activate or suppress) the ability of the antigen recipient to develop either humoral or cytotoxic immune response. On the basis of short peptide activating motifs that have been discovered to affect the immune system, we have developed a family of short synthetic peptides with immunomodulating properties.
These artificial peptides, with a maximum size of eight amino acids, have demonstrated the properties of both local and central action adjuvants,thus affecting the general immunocompetence of the recipient as well as promoting immune response at the site of injection. One of them, named IMMACCEL-R™, showed advantageous properties and excellent performance in accelerating immune response.
Quality of sera prepared using IMMACCEL-R in 28-day immunization Scheme: Immunohistochemistry analysis of formalin-fixed sections of mouse mammary gland using antibodies to whey acidic protein (left) and mouse cytokeratin-18 (right). Immunoblotting analysis of mouse mammary tumour cells using antibodies to mouse cytokeratin-18 and mouse cytokeratin-19.
IMMACCEL-R, when injected into the recipient animal, stimulates induction of cytokine cascades that closely resemble those seen at the early stages of inflammatory response, thus making the animal more prone to give a strong immune response when injected with the antigen.Among the genes affected are those encoding main cytokines and their receptors, as well as several chemokines typical of the inflammatory response . When mixed with antigen and delivered together at the site of injection (intradermal or subcutaneous), IMMACCEL-R further improves the immune response by enhancing presentation of processed antigen by dendritic cells (data not shown) and increasing the specific proliferation of T-helper or T-killer cells, depending on theantigen. Owing to IMMACCEL-R’s effect on immune response, earlier and stronger antibody responses are observed, as well as earlier switch of specific antibody from immunoglobulin M (IgM) to IgG1 and IgG2 isotypes. By day 28, the titer of specific antibodies is already up to 30 times higher for IgGs of γ1 and γ2a isotypes, and 2–4 times lower for IgM. The adjuvant works equally well for all mammalian species tested (mouse, rabbit, goat and swine). A slightly changed version of the molecule is also effective in birds (chicken). The adjuvant is nontoxic and works with all tested types of antigens: proteins, glycoproteins, peptides and haptens conjugated to a carrier.
mRNA expression for different cytokines, as detected by RT-PCR.Mice were injected with either antigen (KLH; blue) or IMMACCEL-R without any antigen (orange); 48 hours later the mRNA profiles for cytokines expressed in spleens were compared to a control untreated mouse of the same sex and age.Data presented as multiples of the respective mRNA levels relative to those in the control sample (spleen of unimmunized mouse). A strong similarity is observed between expression profiles for immunized and IMMACCEL-R–treated naive mouse.
The relative titer of specific antibodies form immunized mice was analyzed by ELISA for samples from day 28. Data are shown as mean + s.d.; three mice in every group. Blue line: control immunization, Orange: IMMACCEL-R.
Immunization with IMMACCEL-R achieves, within 28 days, specific-antibody IgG titers in rabbits similar to those obtained with a standard 80-day protocol. When prepared against strong peptide antigen, the affinity-purified antibodies can be used in many applications and do not differ in affinity or specificity from those obtained through a standard immunization scheme in 80-90 days.
accelerate humoral response
ADJUVANT T1/T4 DATA SHEET
ADJUVANT T1/T4 MSDS
ADJUVANT T1/T4 SAFETY FOR ANIMALS INFORMATION
Groups of 3 rabbits were immunized with antigen (DNP-KLH) with (Orange) or without (Blue) IMMACCEL-R. Serum samples were collected at days 14, 21,28, 35 and analyzed for the presence of specific IgGs to DNP. Shown comparative serum activity at dilution 1:10000 for various time points. Filled area—background reaction