hemolytic plaque technique


Summary: A method to identify and enumerate cells that are synthesizing ANTIBODIES against ANTIGENS or HAPTENS conjugated to sheep RED BLOOD CELLS. The sheep red blood cells surrounding cells secreting antibody are lysed by added COMPLEMENT producing a clear zone of HEMOLYSIS. (From Illustrated Dictionary of Immunology, 3rd ed)

Top Publications

  1. Lee K, Johnson V, Blakley B. The effect of exposure to a commercial 2,4-D formulation during gestation on the immune response in CD-1 mice. Toxicology. 2001;165:39-49 pubmed
    ..Therefore, the impact on human and animal health from an immune perspective, which would be encountered following normal application in the environment, would be minimal. ..
  2. Kastner R, Dussurget O, Archambaud C, Kernbauer E, Soulat D, Cossart P, et al. LipA, a tyrosine and lipid phosphatase involved in the virulence of Listeria monocytogenes. Infect Immun. 2011;79:2489-98 pubmed publisher
    ..Bacteria lacking LipA are severely attenuated in virulence in vivo, thus revealing a so-far-undescribed enzymatic activity involved in Listeria infection...
  3. Ritter D, Kuhn R. Antigen-specific T-helper cells abrogate suppression in Trypanosoma cruzi-infected mice. Infect Immun. 1990;58:3248-56 pubmed
    ..It can be concluded from these data that antigen-specific Th cells can potentiate immune responses in mice infected with T. cruzi but that highly active suppressor macrophages can inhibit the expression of these primed Th cells. ..
  4. Sajid M, Iqbal Z, Muhammad G, Sandhu M, Khan M, Saqib M, et al. Effect of ivermectin on the cellular and humoral immune responses of rabbits. Life Sci. 2007;80:1966-70 pubmed
    ..A graded dose immune response suggested an immunopotentiating effect of ivermectin at higher doses. ..
  5. Wilson S, Munson A, Meade B. Assessment of the functional integrity of the humoral immune response: the plaque-forming cell assay and the enzyme-linked immunosorbent assay. Methods. 1999;19:3-7 pubmed
    ..Both approaches will provide a more extensive delineation of the safety profile of a drug or chemical. The latter approach will also reduce the number of animals needed and the cost of the study. ..
  6. Douglas G, Rubin A. Partial characterization of a T cell-derived factor that suppresses the initiation of the humoral immune response in vitro. Immunology. 1977;33:745-55 pubmed
    ..The antibody initiation suppressor factor (AISF) was concentrated and partially purified by gel filtration, followed by poly-acrylamide gel electrophoresis. ..
  7. Bagg J, Perry K, Parry J, Mortimer P, Peters T. The influence of dental status on the detection of IgG class anti-viral antibodies in human saliva. Arch Oral Biol. 1991;36:221-6 pubmed
    ..The values for sensitivity, specificity and positive predictive value suggest that assay of saliva for antiviral IgG antibody is a satisfactory technique regardless of dental status. ..
  8. van Houte A, Snippe H, Schmitz M, Willers J. Characterization of immunogenic properties of haptenated liposomal model membranes in mice. V. Effect of membrane composition on humoral and cellular immunogenicity. Immunology. 1981;44:561-8 pubmed
  9. Owen F. A mature T lymphocyte marker closely linked to Igh-1 that is expressed on the precursor for the suppressor T cell regulating a primary response to SRBC. J Immunol. 1980;124:1411-7 pubmed
    ..This determinant is expressed on Ts cells for many antigens and may represent a heavy chain linked "constant" portion of the antigen-specific receptor. ..

More Information


  1. Pujol C, Feledi C, Massouh E. Coupling of foot-and-mouth disease virus to sheep red blood cells using tannic acid for immunological assays. J Virol Methods. 1988;20:275-84 pubmed
    ..6. Specificity of the reaction was confirmed by PH and PHI in homologous and heterologous systems. The coupled antigen-antibody complex was sensitive to complement mediated lysis in a PIL test. ..
  2. Hirst R, Yesilkaya H, Clitheroe E, Rutman A, Dufty N, Mitchell T, et al. Sensitivities of human monocytes and epithelial cells to pneumolysin are different. Infect Immun. 2002;70:1017-22 pubmed
    ..Analysis of pneumolysin-treated THP-1 cells by electron microscopy revealed membrane lesions of between 100 and 200 nm in diameter. ..
  3. Arkoosh M, Clemons E, Myers M, Casillas E. Suppression of B-cell mediated immunity in juvenile chinook salmon (Oncorhynchus tshawytscha) after exposure to either a polycyclic aromatic hydrocarbon or to polychlorinated biphenyls. Immunopharmacol Immunotoxicol. 1994;16:293-314 pubmed
  4. Jackson J, Cross R, Walker R, Markesbery W, Brooks W, Roszman T. Influence of serotonin on the immune response. Immunology. 1985;54:505-12 pubmed
    ..Thus, immunomodulation by serotonin appears to be mediated via peripheral mechanism(s). ..
  5. Aguayo Mazzucato C, van Haaren M, Mruk M, Lee T, Crawford C, Hollister Lock J, et al. ? Cell Aging Markers Have Heterogeneous Distribution and Are Induced by Insulin Resistance. Cell Metab. 2017;25:898-910.e5 pubmed publisher
    ..These novel findings about ? cell and islet heterogeneity, and how they change with age, open up an entirely new set of questions about the pathogenesis of type 2 diabetes. ..
  6. Emara A, Draz E. Immunotoxicological study of one of the most common over-the-counter pyrethroid insecticide products in Egypt. Inhal Toxicol. 2007;19:997-1009 pubmed
    ..The present study findings indicate that repeated noncontinuous inhalation of a commonly utilized insecticide that contains imiprothrin and deltamethrin can cause a variety of immunotoxic effects in sites distal to the lungs. ..
  7. DeHeer D, Edgington T. Identification of derepressed autoimmunocompetent B lymphoid cells in NZB mice. Clin Exp Immunol. 1974;16:431-43 pubmed
  8. Doenhoff M, Leuchars E, Kerbel R, Wallis V, Davies A. Adult and pre-adult thymectomy of mice: contrasting effects on immune responsiveness, and on numbers of mitogen-responsive and Thy-1+ lymphocytes. Immunology. 1979;37:397-405 pubmed
    ..The results are discussed with respect to other work on the effects of thymectomy of mice during the post-neonatal and pre-adult period. ..
  9. Boockfor F, Fidan M. Reverse hemolytic plaque assays: versatility in the study of secretion. Methods. 2004;33:273-80 pubmed
    ..Moreover, with the many adaptations possible, it should be particularly valuable in the future for the study of peptide release in other cell types such as neurons. ..
  10. Janssen G, Penninks A, Knippels L, van Zijverden M, Spanhaak S. The evaluation of the immunomodulating properties of ERA-63 a pharmaceutical with estrogenic activity. Toxicol Lett. 2008;180:196-201 pubmed publisher
    ..e. decreases in specific antibody responses and an increased susceptibility for Listeria monocytogenes infects) for both compounds, this indicates that available clinical data for EE facilitates the human risk assessment of ERA-63. ..
  11. Jayakumar K, Honnegowda -, Krishnappa G, Sastry K, Narayana K. Effect of ciprofloxacin on specific immune response in rabbits. Indian J Exp Biol. 2002;40:111-4 pubmed
    ..It can be concluded that ciprofloxacin at the dose and duration employed did not adversely affect specific immune response in normal rabbits. ..
  12. Iwasaki T, Hamano T, Aizawa K, Kobayashi K, Kakishita E. Characterization of a functional null cell line derived from NZB x NZW)F1 mice. Scand J Immunol. 1994;40:669-74 pubmed
  13. Nossal G, Lewis H. Functional symmetry amongst daughter cells arising in vitro from single antibody-forming cells. Immunology. 1971;20:739-53 pubmed
  14. Baecher Steppan L, Kerkvliet N. The haemolytic antibody isotope release (HAIR) assay: an efficient alternative technique to conventional plaque assays. Clin Exp Immunol. 1981;44:440-4 pubmed
    ..The HAIR assay is an easy, sensitive, and reproducible technique that is especially useful when large numbers of animals are required for testing. ..
  15. Paramonov B, Klimova O, Chebotarev V. Effects of collagenolytic protease preparations from invertebrates on the immune system. Bull Exp Biol Med. 2000;129:547-9 pubmed
    ..These substances in therapeutic doses did not affect the humoral immune response estimated by hemagglutination and local hemolysis in gel and caused no local irritation and allergic reactions after long-term application to the skin. ..
  16. Ratajczak H, Sothern R, Dozier M, Gaworski C. Immunomodulatory screening test of corn oil administered orally to female mice: effect of timing of dosing within 24 hours. Chronobiol Int. 1999;16:185-97 pubmed
  17. Choi U, Lee S, Kim K, Kim D, Choi K, Cha S, et al. Is a booster dose necessary in children after immunization with live attenuated Japanese encephalitis vaccine?. J Trop Pediatr. 2013;59:423-5 pubmed publisher
    ..Thus, we predict that immunity will be maintained for a long time by an amnestic response. Low percentages of adverse reactions indicated the safety of the immunizations. ..
  18. Martens R, Cohen N, Chaffin M, Takai S, Doherty C, Angulo A, et al. Evaluation of 5 serologic assays to detect Rhodococcus equi pneumonia in foals. J Am Vet Med Assoc. 2002;221:825-33 pubmed
    ..Results suggest that serologic assays, whether performed on single or paired samples, cannot be used to reliably establish, confirm, or exclude a diagnosis of R equi pneumonia in foals. ..
  19. Koros A, Hamill E. Mechanisms of suppression of the immune response. I. Differences in the effect of specific inhibitory antibody on distribution of 51CR-labelled sheep erythrocytes in different mouse strains. Immunology. 1973;25:717-36 pubmed
  20. McCracken A, McBride W, Weir D. Adjuvant-induced antired blood cell activity in CBA mice. Clin Exp Immunol. 1971;8:949-55 pubmed
  21. Hutchings P, Varey A, Cooke A. Immunological defects in SJL mice. Immunology. 1986;59:445-50 pubmed
    ..DBA mice are also shown, in this report, to have small numbers of antibody-forming cells to bromelain-treated autologous red blood cells but to be capable of responding to LPS in vivo with an increase in SRBC and TNP antibody responses. ..
  22. Sohn Y, Tandan J, Yoksan S, Ji M, Ohrr H. A 5-year follow-up of antibody response in children vaccinated with single dose of live attenuated SA14-14-2 Japanese encephalitis vaccine: immunogenicity and anamnestic responses. Vaccine. 2008;26:1638-43 pubmed publisher
    ..However, further studies should be carried out to support whether one dose of live JE vaccine is sufficient for people in JE non-endemic area. ..
  23. Lundeberg T, Eriksson S, Theodorsson E. Neuroimmunomodulatory effects of acupuncture in mice. Neurosci Lett. 1991;128:161-4 pubmed
    ..These results suggest that acupuncture, by activation of the autonomic nervous system, modulates the immune response. ..
  24. Malee M, Wu K. Corticosteroid dynamics in the nonpregnant, pregnant, and postpartum spontaneously hypertensive rat. Am J Hypertens. 2000;13:410-7 pubmed
  25. Ahmad A, Mond J. Restoration of in vitro responsiveness of xid B cells to TNP-Ficoll by 8-mercaptoguanosine. J Immunol. 1986;136:1223-6 pubmed
  26. Sprent J, Bruce J. Physiology of B cells in mice with X-linked immunodeficiency (xid). III. Disappearance of xid B cells in double bone marrow chimeras. J Exp Med. 1984;160:711-23 pubmed
    ..The finding that xid B cells eventually disappear in the presence of normal B cells strengthens the view that xid B cells are an abnormal population not represented in normal mice. ..
  27. Johnson S. A B-cell hybridoma product inhibiting antibody secretion via 5'-nucleotidase. Clin Exp Immunol. 1985;62:412-20 pubmed
    ..The factor was called BAN (B cell anti 5'-nucleotidase). The suppressive action of BAN on IgG production was blocked by antibodies against 5'-nucleotidase suggesting that the lymphocyte enzyme may be acting as a BAN receptor. ..
  28. Pepeljnjak S, Zorc B, Butula I. Antimicrobial activity of some hydroxamic acids. Acta Pharm. 2005;55:401-8 pubmed
    ..Compound showed no antimicrobial activity on any of the tested microorganisms. Other compounds showed a narrow spectrum of antibacterial activity, but no antifungal activity. ..
  29. Wieczorek Z, Słoń J, Kluczyk A, Zbozień R, Stafanowicz P, Siemion I. The immunomodulatory diversity of the proteins of the transforming growth factor beta (TGF beta) family. Int J Pept Protein Res. 1995;46:113-8 pubmed
    ..The results obtained also suggest that chicken TGF beta 4 should be associated with immunostimulative effects and xenopus TGF beta 5 with immunosuppressive ones. ..
  30. Chetty K, Brown K, Walker J, Woods L. Effects of chlordecone and malnutrition on immune response in rats. Life Sci. 1993;52:PL175-80 pubmed
    ..Similar increase in PFC was observed in rats fed on Ca-D but not P-D diet containing 10 or 100 ppm of Cd. Whereas, rats fed on Ca+P-D diet containing 100 ppm of Cd exhibited a significant decrease in PFC. ..
  31. Tingle A, Yang T, Allen M, Kettyls G, Larke R, Schulzer M. Prospective immunological assessment of arthritis induced by rubella vaccine. Infect Immun. 1983;40:22-8 pubmed
    ..Rubella lymphoproliferative responses were elevated at 6 weeks post-immunization in the group developing arthralgia or arthritis or both, with no difference between the groups observed at 6 months post-immunization. ..
  32. Bittencourt C, Azzolini A, Ferreira D, Assis Pandochi A. Antibody responses in hyperthyroid rats. Int Immunopharmacol. 2007;7:989-93 pubmed
    ..It is suggested that antibody production is affected by thyroid hormone levels. ..
  33. Devoino L, Cheido M, Al perina E. Involvement of the nucleus accumbens in stimulation of the immune response in rats after activation of opioid mu receptors with DAGO. Neurosci Behav Physiol. 2002;32:529-32 pubmed
    ..These data provide evidence for the involvement of the nucleus accumbens in the process of immunomodulation and for the importance of opioid mu receptors in the nucleus accumbens in the stimulation of immunogenesis. ..
  34. Sopori M, Kozak W, Savage S, Geng Y, Soszynski D, Kluger M, et al. Effect of nicotine on the immune system: possible regulation of immune responses by central and peripheral mechanisms. Psychoneuroendocrinology. 1998;23:189-204 pubmed
    ..These results support the hypothesis that NT alters immune responses by directly interacting with T cells, as well as indirectly through brain-immune interactions. ..
  35. Chen L, Wang Y, He S, Liao G, Zhang Z. [Study on the toxic reaction induced by single dose of qingkailing injection]. Zhong Yao Cai. 2011;34:254-8 pubmed
    ..In clinical practice some adverse reactions induced by Qingkailing injection occurred concomitantly with acute hemolysis within vessels, which might be caused by honeysuckle and gardenia. And the hemolysis was independent of allergy. ..
  36. Harris G. Antibody production in vitro. The effects of anti-allotypic serum on the secondary response of rabbit spleen cell suspensions to sheep erythrocytes (SRC). Immunology. 1969;17:911-26 pubmed
  37. Khajuria A, Gupta A, Garai S, Wakhloo B. Immunomodulatory effects of two sapogenins 1 and 2 isolated from Luffa cylindrica in Balb/C mice. Bioorg Med Chem Lett. 2007;17:1608-12 pubmed
    ..Overall, sapogenins 1 and 2 showed dose relative immunostimulatory effect on in vivo immune functions in mice. ..
  38. Lee J, Hampson D, Lymbery A, Harders S. The porcine intestinal spirochaetes: identification of new genetic groups. Vet Microbiol. 1993;34:273-85 pubmed
    ..We consider that group C represents a new genus of spirochaetes, members of which may be associated with spirochaetal diarrhoea...
  39. Glavaski Joksimovic A, Jeftinija K, Jeremic A, Anderson L, Jeftinija S. Mechanism of action of the growth hormone secretagogue, L-692,585, on isolated porcine somatotropes. J Endocrinol. 2002;175:625-36 pubmed
  40. Smith H, Yaffe L, Kastner D, Steinberg A. Evidence that Lyb-5 is a differentiation antigen in normal and xid mice. J Immunol. 1986;136:1194-200 pubmed
    ..Some generation of Lyb-5+ antibody-forming cells occurred in the last 24 hr of culture. These studies suggest that the Lyb-5 marker represents a differentiation antigen on relatively mature B cells. ..
  41. Smialowicz R, Burgin D, Williams W, Diliberto J, Setzer R, Birnbaum L. CYP1A2 is not required for 2,3,7,8-tetrachlorodibenzo-p-dioxin-induced immunosuppression. Toxicology. 2004;197:15-22 pubmed
    ..Thus, CYP1A2 is not required for TCDD-induced immunosuppression in the mouse. ..
  42. Wu C, Bey R, Loken K. Purification and characterization of the 1q subcomponent of canine complement and its use in the 125I-C1q binding assay for detection of immune complexes. Am J Vet Res. 1988;49:865-9 pubmed
    ..Isolated canine C1q was iodinated, and the fluid-phase binding assay was used to detect circulating immune complexes in dogs with systemic lupus erythematosus and rheumatoid arthritis. ..
  43. Hunter P, Kettman J. Mode of action of a supernatant activity from T-cell cultures that nonspecifically stimulates the humoral immune response. Proc Natl Acad Sci U S A. 1974;71:512-6 pubmed
  44. Shultz L, Green M. Motheaten, an immunodeficient mutant of the mouse. II. Depressed immune competence and elevated serum immunoglobulins. J Immunol. 1976;116:936-43 pubmed
    ..Motheaten mice, thus, appear to have a severe immune deficiency, but the basic nature of the deficiency is not yet known. ..
  45. Babcook J, Leslie K, Olsen O, Salmon R, Schrader J. A novel strategy for generating monoclonal antibodies from single, isolated lymphocytes producing antibodies of defined specificities. Proc Natl Acad Sci U S A. 1996;93:7843-8 pubmed
  46. Devoino L, Cheido M, Al perina E. Involvement of the rat caudate nucleus in the immunostimulatory effect of DAGO. Neurosci Behav Physiol. 2001;31:323-6 pubmed
    ..Since the effect of immunostimulation induced by DAGO disappeared when given to animals with caudate nucleus lesions, it was concluded that this structure is involved in activatory immunogenesis via mu opioid mechanisms. ..
  47. Vila Porcile E, Barret A, Corvol P. Secretion of renin-angiotensin system (RAS) components by normal and tumoral lactotropes. A comparative study using reverse hemolytic plaque assay (RHPA) and immunoelectron microscopy. J Histochem Cytochem. 2000;48:1691-704 pubmed
    ..The cleaved components in the RER suggest the existence of early maturation, whereas the presence of uncleaved products in the secretory granules of normal lactotropes might indicate late maturation sites. ..
  48. Murunikkara V, Rasool M. Trikatu, an herbal compound as immunomodulatory and anti-inflammatory agent in the treatment of rheumatoid arthritis--an experimental study. Cell Immunol. 2014;287:62-8 pubmed publisher
    ..Thus, in conclusion, our data suggest that trikatu could be considered as a potential anti-inflammatory agent for treating autoimmune inflammatory disorders like rheumatoid arthritis with immunosuppressive property. ..
  49. Lehner P, Hutchings P, Lydyard P, Cooke A. II. IgM-mediated enhancement: dependency on antigen dose, T-cell requirement and lack of evidence for an idiotype-related mechanism. Immunology. 1983;50:503-9 pubmed
    ..The IgM enhancement phenomenon has been demonstrated in several strains of mice, providing strong evidence against an idiotype-related mechanism. ..
  50. Song X, Li C, Zeng Y, Wu H, Huang Z, Zhang J, et al. Immunomodulatory effects of crude phenylethanoid glycosides from Ligustrum purpurascens. J Ethnopharmacol. 2012;144:584-91 pubmed publisher
    ..The results suggested that phenylethanoid glycosides from Ligustrum purpurascens have immunomodulatory effects on mice. ..
  51. Zaidi S, Raisuddin S, Singh K, Jafri A, Husain R, Husain M, et al. Acrylamide induced immunosuppression in rats and its modulation by 6-MFA, an interferon inducer. Immunopharmacol Immunotoxicol. 1994;16:247-60 pubmed
    ..Potentiation of EAC-rosettes, hemagglutination titre, IgM-PFC and DTH response against SRBC was observed. It is concluded that 6-MFA ameliorate the ACR induced toxicity. This study may be of significance in prevention of ACR toxicity. ..
  52. Escamilla Chimal E, Hiriart M, Sánchez Soto M, Fanjul Moles M. Serotonin modulation of CHH secretion by isolated cells of the crayfish retina and optic lobe. Gen Comp Endocrinol. 2002;125:283-90 pubmed
    ..The authors conclude that (1) retinal CHH-secreting cells correspond to a population of retinal tapetal cells and (2) optic lobe CHH-secreting cells correspond to two subpopulations of CHH of medulla terminalis-X organ. ..
  53. Chotpitayasunondh T, Sohn Y, Yoksan S, Min J, Ohrr H. Immunizing children aged 9 to 15 months with live attenuated SA14-14-2 Japanese encephalitis vaccine in Thailand. J Med Assoc Thai. 2011;94 Suppl 3:S195-203 pubmed
    ..Nevertheless, its long-term immune response and possible influences from natural dengue infection requires further evaluation. ..