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Chapter 37 : Functional Assays for B Cells and Antibodies
Authors:
Moon H. Nahm,
Robin G. Lorenz
Content Type: Reference
Publication Year:
2006
Category: Immunology
Book DOI:
10.1128/9781555815905
Chapter DOI:
10.1128/9781555815905.ch37
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Functional Assays for B Cells and Antibodies, Page 1 of 2
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Abstract:
The function of the humoral immune response differs depending on the class of antibody produced and the differentiation state of the B cell. Primary defects in the humoral components of the immune response are usually recognized by clues as frequent development of infections early in life and difficulty in clearance of infections. The most direct measure of in vivo B-cell function is immunoglobulin secretion by plasma cells. Serum immunoglobulins are most commonly measured by automated nephelometry. Antibodies provide protection to the host in various ways. Antibodies can neutralize toxins (e.g., tetanus toxin), neutralize viruses, prevent the adhesion of bacteria to the host cells, kill bacteria in the presence of complement, and opsonize bacteria for phagocytes. Generally, the complement-mediated bactericidal mechanism does not kill gram-positive bacteria, although there are exceptional cases. This mechanism is primarily relevant in the study of antibodies to gram-negative bacteria, which have thin walls. Opsonophagocytosis is the primary protective mechanism of antibodies against gram-positive bacteria. Various methods have been developed to measure the opsonizing capacity of antibodies in vitro. The classical approach is to perform an opsonophagocytic killing assay. In this assay, the bacteria are opsonized (coated) with antibodies and complement. The increased use of functional assays has shown the importance of standardization.
Citation:
Nahm M, Lorenz R.
2006.
Functional Assays for B Cells and Antibodies,
p 328-336.
In
Detrick B, Hamilton R, Folds J (ed),
Manual of Molecular and Clinical Laboratory Immunology, 7th Edition.
ASM Press, Washington, DC.
doi: 10.1128/9781555815905.ch37
Key Concept Ranking
- Memory B Cell
- 0.62759674
- B Cells
- 0.54813975
- Adaptive Immune System
- 0.5136671
- Innate Immune System
- 0.5136671
- Humoral Immune Response
- 0.4430558
0.62759674
Figures
Functional Assays for B Cells and Antibodies
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Citation:
Nahm M, Lorenz R.
2006.
Functional Assays for B Cells and Antibodies,
p 328-336.
In
Detrick B, Hamilton R, Folds J (ed),
Manual of Molecular and Clinical Laboratory Immunology, 7th Edition.
ASM Press, Washington, DC.
doi: 10.1128/9781555815905.ch37
/content/10.1128/9781555815905.ch37.ch37fig01
FIGURE 1
Setup template for assay with multiple plates. (A) First plate arrangement. (B) Additional plate arrangement.
10.1128/9781555815905/f0334-01_thmb.gif
10.1128/9781555815905/f0334-01.gif
FIGURE 1
Setup template for assay with multiple plates. (A) First plate arrangement. (B) Additional plate arrangement.
Citation:
Nahm M, Lorenz R.
2006.
Functional Assays for B Cells and Antibodies,
p 328-336.
In
Detrick B, Hamilton R, Folds J (ed),
Manual of Molecular and Clinical Laboratory Immunology, 7th Edition.
ASM Press, Washington, DC.
doi: 10.1128/9781555815905.ch37
References
/content/book/10.1128/9781555815905.ch37
1.
Agematsu, K.,, T. Futatani,, S. Hokibara,, N. Kobayashi,, M. Takamoto,, S. Tsukada,, H. Suzuki,, S. Koyasu,, T. Miyawaki,, K. Sugane,, A. Komiyama, and , H. D. Ochs. 2002. Absence of memory B cells in patients with common variable immunodeficiency. Clin. Immunol. 103: 34– 42.
2.
Aguilar, P.,, E. Renoult,, L. Jarrosson,, M. N. Kolopp-Sarda,, C. P. Mathieu,, G. C. Faure,, M. Kessler,, M. C. Bene,, C. Kohler, and , A. Kennel De March. 2003. Anti-HBs cellular immune response in kidney recipients before and 4 months after transplantation. Clin. Diagn. Lab. Immunol. 10: 1117– 1122.
3.
Alam, R., and , M. Gorska. 2003. 3. Lymphocytes. J. Allergy Clin. Immunol. 111: S476– S485.
4.
Balmer, P.,, J. North,, D. Baxter,, E. Stanford,, A. Melegaro,, E. B. Kaczmarski,, E. Miller, and , R. Borrow. 2003. Measurement and interpretation of pneumococcal IgG levels for clinical management. Clin. Exp. Immunol. 133: 364– 369.
5.
Bergeron, M. G.,, P. Simard, and , P. J. Provencher. 1987. Influence of growth medium and supplement on growth of Haemophilus influenzae and on antibacterial activity of several antibiotics. J. Clin. Microbiol. 25: 650– 655.
6.
Bogaert, D.,, M. Sluijter,, P. De Groot, and , R. W. Hermans. 2004. Multiplex opsonophagocytosis assay (MOPA): a useful tool for the monitoring of the 7-valent pneumococcal conjugate vaccine. Vaccine 22: 4014– 4020.
7.
Buckley, R. H. 2002. Immunoglobulin G subclass deficiency: fact or fancy? Curr. Allergy Asthma Rep. 2: 356– 360.
8.
Buckley, R. H.,, S. C. Dees, and , W. M. O’Fallon. 1968. Serum immunoglobulins. I. Levels in normal children and in uncomplicated childhood allergy. Pediatrics 41: 600– 611.
9.
Butch, A. W.,, K. A. Macke,, M. G. Scott,, M. Inkster, and , M. H. Nahm. 1989. Mitogen-induced human IgG subclass expression. II. IgG1 and IgG3 subclasses are preferentially stimulated by a combination of Staphylococcus aureus Cowan I and pokeweed mitogen. Hum. Immunol. 24: 207– 218.
10.
Butch, A. W., and , M. H. Nahm. 1992. Functional properties of human germinal center B cells. Cell. Immunol. 140: 331– 344.
11.
Carneiro-Sampaio, M. M.,, A. S. Grumach, and , A. Manissadjian. 1991. Laboratory screening for the diagnosis of children with primary immunodeficiencies. J. Investig. Allergol. Clin. Immunol. 1: 195– 200.
12.
Fleck, R. A.,, S. Romero-Steiner, and , M. H. Nahm. 2005. Use of HL-60 cell line to measure opsonic capacity of pneumococcal antibodies. Clin. Diagn. Lab. Immunol. 12: 19– 27.
13.
Fleisher, T. A., and , J. B. Oliveira. 2004. Functional and molecular evaluation of lymphocytes. J. Allergy Clin. Immunol. 114: 227-234; quiz 235.
14.
Folds, J. D., and , J. L. Schmitz. 2003. 24. Clinical and laboratory assessment of immunity. J. Allergy Clin. Immunol. 111: S702– S711.
15.
Fusco, P. C.,, J. W. Perry,, S. M. Liang,, M. S. Blake,, F. Michon, and , J. Y. Tai. 1997. Bactericidal activity elicited by the beta C protein of group B streptococci contrasted with capsular polysaccharides. Adv. Exp. Med. Biol. 418: 841– 845.
16.
James, S. P. 2004. Measurement of basic immunologic characteristics of human mononuclear cells, p. 7.10.1– 7.10.6. In J. E. Coligan,, A. M. Kruisgeek,, D. H. Margulies,, E. M. Shevach, and , W. Strober (ed.), Current Protocols in Immunology. John Wiley & Sons, New York, N.Y.
17.
Jansen, W. T.,, M. Vakevainen-Anttila,, H. Kayhty,, M. Nahm,, N. Bakker,, J. Verhoef,, H. Snippe, and , A. F. Verheul. 2001. Comparison of a classical phagocytosis assay and a flow cytometry assay for assessment of the phagocytic capacity of sera from adults vaccinated with a pneumococcal conjugate vaccine. Clin. Diagn. Lab. Immunol. 8: 245– 250.
18.
Kim, K. H.,, J. Yu, and , M. H. Nahm. 2003. Efficiency of a pneumococcal opsonophagocytic killing assay improved by multiplexing and by coloring colonies. Clin. Diagn. Lab. Immunol. 10: 616– 621.
19.
Langermann, S.,, S. Palaszynski,, M. Barnhart,, G. Auguste,, J. S. Pinkner,, J. Burlein,, P. Barren,, S. Koenig,, S. Leath,, C. H. Jones, and , S. J. Hultgren. 1997. Prevention of mucosal Escherichia coli infection by FimH-adhesin-based systemic vaccination. Science 276: 607– 611.
20.
Lin, J. S.,, M. K. Park, and , M. H. Nahm. 2001. Chromogenic assay measuring opsonophagocytic killing capacities of antipneumococcal antisera. Clin. Diagn. Lab. Immunol. 8: 528– 533.
21.
Maguire, G. A.,, D. S. Kumararatne, and , H. J. Joyce. 2002. Are there any clinical indications for measuring IgG subclasses? Ann. Clin. Biochem. 39: 374– 377.
22.
Martin, F., and , J. F. Kearney. 2001. B1 cells: similarities and differences with other B cell subsets. Curr. Opin. Immunol. 13: 195– 201.
23.
Martinez, J.,, T. Pilishvili,, S. Barnard,, J. Caba,, W. Spear,, S. Romero-Steiner, and , G. M. Carlone. 2002. Opsonophagocytosis of fluorescent polystyrene beads coupled to Neisseria meningitidis serogroup A, C, Y, or W135 polysaccharide correlates with serum bactericidal activity. Clin. Diagn. Lab. Immunol. 9: 485– 488.
24.
Martinez, J. E.,, S. Romero-Steiner,, T. Pilishvili,, S. Barnard,, J. Schinsky,, D. Goldblatt, and , G. M. Carlone. 1999. A flow cytometric opsonophagocytic assay for measurement of functional antibodies elicited after vaccination with the 23-valent pneumococcal polysaccharide vaccine. Clin. Diagn. Lab. Immunol. 6: 581– 586.
25.
McHeyzer-Williams, M. G. 2003. B cells as effectors. Curr. Opin. Immunol. 15: 354– 361.
26.
Mountzouros, K. T., and , A. P. Howell. 2000. Detection of complement-mediated antibody-dependent bactericidal activity in a fluorescence-based serum bactericidal assay for group B Neisseria meningitidis. J. Clin. Microbiol. 38: 2878– 2884.
27.
Nahm, M. H.,, R. M. Blaese,, M. J. Crain, and , D. E. Briles. 1986. Patients with Wiskott-Aldrich syndrome have normal IgG2 levels. J. Immunol. 137: 3484– 3487.
28.
Nahm, M. H.,, D. E. Briles, and , X. Yu. 2000. Development of a multi-specificity opsonophagocytic killing assay. Vaccine 18: 2768– 2771.
29.
Noroski, L. M., and , W. T. Shearer. 1998. Screening for primary immunodeficiencies in the clinical immunology laboratory. Clin. Immunol. Immunopathol. 86: 237– 245.
30.
Piqueras, B.,, C. Lavenu-Bombled,, L. Galicier,, F. Bergeron-van der Cruyssen,, L. Mouthon,, S. Chevret,, P. Debre,, C. Schmitt, and , E. Oksenhendler. 2003. Common variable immunodeficiency patient classification based on impaired B cell memory differentiation correlates with clinical aspects. J. Clin. Immunol. 23: 385– 400.
31.
Rodriguez, T.,, M. Lastre,, B. Cedre,, J. del Campo,, G. Bracho,, C. Zayas,, C. Taboada,, M. Diaz,, G. Sierra, and , O. Perez. 2002. Standardization of Neisseria meningitidis serogroup B colorimetric serum bactericidal assay. Clin. Diagn. Lab. Immunol. 9: 109– 114.
32.
Romero-Steiner, S.,, J. Fernandez,, C. Biltoft,, M. E. Wohl,, J. Sanchez,, J. Feris,, S. Baiter,, O. S. Levine, and , G. M. Carlone. 2001. Functional antibody activity elicited by fractional doses of Haemophilus influenzae type b conjugate vaccine (polyribosylribitol phosphate-tetanus toxoid conjugate). Clin. Diagn. Lab. Immunol. 8: 1115– 1119.
33.
Romero-Steiner, S.,, C. Frasch,, N. Concepcion,, D. Goldblatt,, H. Kayhty,, M. Vakevainen,, C. Laferriere,, D. Wauters,, M. H. Nahm,, M. F. Schinsky,, B. D. Plikaytis, and , G. M. Carlone. 2003. Multilaboratory evaluation of a viability assay for measurement of opsonophagocytic antibodies specific to the capsular polysaccharides of Streptococcus pneumoniae. Clin. Diagn. Lab. Immunol. 10: 1019– 1024.
34.
Romero-Steiner, S.,, W. Spear,, N. Brown,, P. Holder,, T. Hennessy,, P. Gomez De Leon, and , G. M. Carlone. 2004. Measurement of serum bactericidal activity specific for Haemophilus influenzae type b by using a chromogenic and fluorescent metabolic indicator. Clin. Diagn. Lab. Immunol. 11: 89– 93.
35.
Scott, M. G., and , M. H. Nahm. 1984. Mitogen-induced human IgG subclass expression. J. Immunol. 133: 2454– 2460.
36.
Shearer, W. T.,, H. M. Rosenblatt,, R. S. Gelman,, R. Oyomopito,, S. Plaeger,, E. R. Stiehm,, D. W. Wara,, S. D. Douglas,, K. Luzuriaga,, E. J. McFarland,, R. Yogev,, M. H. Rathore,, W. Levy,, B. L. Graham, and , S. A. Spector. 2003. Lymphocyte subsets in healthy children from birth through 18 years of age: the Pediatric AIDS Clinical Trials Group P1009 study. J. Allergy Clin. Immunol. 112: 973– 980.
37.
Thiel, A.,, A. Scheffold, and , A. Radbruch. 2004. Antigen-specific cytometry—new tools arrived! Clin. Immunol. 111: 155– 161.
38.
Vermont, C., and , G. van den Dobbelsteen. 2002. Neisseria meningitidis serogroup B: laboratory correlates of protection. FEMS Immunol. Med. Microbiol. 34: 89– 96.
39.
Warnatz, K.,, A. Denz,, R. Drager,, M. Braun,, C. Groth,, G. Wolff-Vorbeck,, H. Eibel,, M. Schlesier, and , H. H. Peter. 2002. Severe deficiency of switched memory B cells (CD27(+)IgM(—)IgD(—)) in subgroups of patients with common variable immunodeficiency: a new approach to classify a heterogeneous disease. Blood 99: 1544– 1551.
40.
Weber-Mzell, D.,, P. Kotanko,, A. C. Hauer,, U. Goriup,, J. Haas,, N. Lanner,, W. Erwa,, I. A. Ahmaida,, S. Haitchi-Petnehazy,, M. Stenzel,, G. Lanzer, and , J. Deutsch. 2004. Gender, age and seasonal effects on IgA deficiency: a study of 7293 Caucasians. Eur. J. Clin. Investig. 34: 224– 228.
Tables
Functional Assays for B Cells and Antibodies
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Citation:
Nahm M, Lorenz R.
2006.
Functional Assays for B Cells and Antibodies,
p 328-336.
In
Detrick B, Hamilton R, Folds J (ed),
Manual of Molecular and Clinical Laboratory Immunology, 7th Edition.
ASM Press, Washington, DC.
doi: 10.1128/9781555815905.ch37
/content/10.1128/9781555815905.ch37.ch37tab01
TABLE 1
B-cell markers used in flow cytometry
TABLE 1
B-cell markers used in flow cytometry
Citation:
Nahm M, Lorenz R.
2006.
Functional Assays for B Cells and Antibodies,
p 328-336.
In
Detrick B, Hamilton R, Folds J (ed),
Manual of Molecular and Clinical Laboratory Immunology, 7th Edition.
ASM Press, Washington, DC.
doi: 10.1128/9781555815905.ch37
/content/10.1128/9781555815905.ch37.ch37tab02
TABLE 2
Associations with humoral or B-lymphocyte deficiencies
TABLE 2
Associations with humoral or B-lymphocyte deficiencies
Citation:
Nahm M, Lorenz R.
2006.
Functional Assays for B Cells and Antibodies,
p 328-336.
In
Detrick B, Hamilton R, Folds J (ed),
Manual of Molecular and Clinical Laboratory Immunology, 7th Edition.
ASM Press, Washington, DC.
doi: 10.1128/9781555815905.ch37
/content/10.1128/9781555815905.ch37.ch37tab03
TABLE 3
In vitro activators for B-cell proliferation and immunoglobulin production
TABLE 3
In vitro activators for B-cell proliferation and immunoglobulin production
Citation:
Nahm M, Lorenz R.
2006.
Functional Assays for B Cells and Antibodies,
p 328-336.
In
Detrick B, Hamilton R, Folds J (ed),
Manual of Molecular and Clinical Laboratory Immunology, 7th Edition.
ASM Press, Washington, DC.
doi: 10.1128/9781555815905.ch37
/content/10.1128/9781555815905.ch37.ch37tab04
TABLE 4
Pneumococcal strains used as targets for the opsonization assay
TABLE 4
Pneumococcal strains used as targets for the opsonization assay
Citation:
Nahm M, Lorenz R.
2006.
Functional Assays for B Cells and Antibodies,
p 328-336.
In
Detrick B, Hamilton R, Folds J (ed),
Manual of Molecular and Clinical Laboratory Immunology, 7th Edition.
ASM Press, Washington, DC.
doi: 10.1128/9781555815905.ch37