Katherine L. Knight, PhD

Katherine L. Knight, PhD

Professor

Ph.D., Indiana University

HOME OF THE RABBIT HYBRIDOMAS

Major Research Interests: B-lymphocyte development and generation of the antibody repertoire

 

Exposure of individuals to foreign antigens, such as those associated with bacteria and viruses, results in production of thousands of different antibody molecules. My laboratory is interested in how organisms develop such a large and diverse array of antibodies during an immune response. The goals of our research are to elucidate the molecular genetic mechanisms that generate antibody diversity and that regulate the expression of immunoglobulin (lg) genes

Mammals use several strategies to generate antibody diversity. They have the capacity to produce many different Ig heavy chains and light chains (the building blocks of antibody molecules), which can be used in different combinations to generate antibody diversity. The Ig heavy and light chains each have variable (V) regions and constant regions that are encoded by separate genes. Different V genes, of which there are hundreds, are used to produce antibodies of different specificities. Rearrangement of the V genes by somatic recombination is required before an antibody can be produced by a B lymphocyte. Additional diversity is generated in gut-associated lymphoid tissues, and exposure to microbial flora of the gut is required for this somatic diversification to occur. We are investigating how a lymphocyte chooses to rearrange and express a particular V gene, how this rearranged V gene undergoes somatic diversification, and how intestinal microorganisms mediate somatic diversification oft heV genes. Details of individual projects can be found at our laboratory home page.

For our investigations we use normal and transgenic rabbits as well as rabbits genetically modified by nuclear transfer techniques. We use molecular genetic, immunochemical, and cell culture techniques, including DNA cloning and sequencing and in vitro gene expression.

Rabbit Hybridomas

 

  Molecular Biology

  • Large Scale Preparation of Plasmid DNA
  • Plasmid DNA Preparation - SuperQuickie
  • Northern Blot Analysis
  • Phage and Phage DNA Preparation
  • Genomic DNA Preparation
  • RNA Isolation by CsCl or TRIzol® Method
  • RNase Protection Assay
  • Preparation of Frozen Electrocompetent E. coli for Transformation
  • Transfection by Electroporation
  • 125I Labeling of Protein - ICl Method
  • Preparation of Single Stranded M13 Template DNA
  • One-step" Protocol for Transforming Yeast
  • Preparation of Rabbit Peripheral Blood Lymphocytes (PBL)

 

Immunology

  • Isolation of PBL from Buffy Coats
  • Western Blot

Mentoring

Mentoring is one of the most important and rewarding experiences of an academic scientist. I have discussed my views on mentoring both in writing and in oral presentations. Five of these are provided below:

• Presentation on Mentoring at the University of Massachusetts, May 2012: Mentoring: The challenge of looking at something familiar and seeing it in a new way

• An article published in the Association for Women in Science - Chicago, February 2010, about my mentoring activities.

• Address at Loyola University Chicago Graduate School Commencement, April 2004 Upon Receipt of Faculty Member of the Year Award.

• Excerpts from the AAI Presidential Address:"Doing it together: A Perspective on the Process of Experimental Science" (1997).

• Comments from Mentoring Symposium in April, 1994 given in a symposium organized by the Women's Committee of AAI at the annual meeting of AAI in 1994.

Kennedy, D. E. and Knight, K.L. (2015) Adipocyte-induced myeloid-derived suppressor cells inhibit B-lymphopoiesis.  J. Immunology, August 12, 2015, doi:10.4049/​jimmunol.1500957

Johnston, P.F., Gerding, D.N., and Knight, K.L.  (2014) Protection from Clostridium difficile infection in CD4 T cell and polymeric immunoglobulin receptor deficient mice.  Infection and Immunity, 82:522-53.  PMID 24218482 “Spotlighted” by the editors

Zhai, S.-K., Volgina, V.V., Sethupathi, P., Knight, K.L., and Lanning, D.  (2014) Chemokine-mediated B cell trafficking during early rabbit GALT development. J. Immunology, 193:5951-5959. PMID: 25385821

Jones, S.E., Paynich, M.L., Kearns, D.B., and Knight, K.L. (2014)  Protection from intestinal inflammation by bacterial exopolysaccharides.  J. Immunology, 192:4813-4820  PMID: 24740503

Jones, S. E., Paynich, M.L., and Knight, K.L. (2014)  Exopolysaccharides:  Sweet success with probiotic therapeutics.  Inflammation and Cell Signaling, 1:e334  doi: 10.14800/ics.334

Semenyuk, E.G., Laning, M.L., Foley, J., Johnston, P.F., Knight, K.L., Gerding, D.N., and Driks A. (2014) Spore formation and toxin production in Clostricuium difficile biofilms.  PLoS One 30;9(1):e87757 PMID:24498186

Yeramilli, V.A., and Knight, K.L., (2013)  Development of CD27+ Marginal zone B cells requires GALT.  Eu. J. Immunology, 43:1484-1488 PMID:23468368

 Jones, S.E. and Knight, K.L. (2012) Bacillus subtilis-mediated protection from Citrobacter rodentium-associated enteric disease requires espH and hagInfection and Immunity, 80:710-719 PMID:22144475 PMC3264298

Bilwani, F. and Knight, K.L., (2012) Adipocyte-derived factor(s) inhibits early stages of B-lymphopoiesis in rabbits and humans, J. Immunology, 189:4379-4386. PMID 23002443

Beaudette-Zlatanova, B.C., Le, P.T., Stiff, P., and Knight, K.L., (2012) A potential role for B cells in suppressed immune responses in cord blood transplant recipients. Bone Marrow Transplantation, 48:85-93  PMID 22732699

Robbins, G. and Knight, K.L.  (2011) Mechanism for Pre-B Cell Loss in VH-Mutant Rabbits, J. Immunol.  187:4714-4720  PMID 21957145 PMC3339493

Siewe, B.T., Kalis, S.L., Le, P.T., Witte, P.L., Choi, S., Conway, S.J., Druschitz, L., and Knight, K.L.  (2011)  In vitro requirement for periostin in B lymphopoiesis Blood 117:3770-3779. PMID 21285437

Siewe, B.T., Kalis, S., Esteves, P. J., Zhou, T., and Knight, K. L. (2010) A novel functional rabbit IL-7 isoform. Developmental and Comparative Immunology, 34:828-36.

Yeramilli, V.A. and Knight, K. L.  (2010)  Requirement for BAFF and APRIL during B cell development in GALT., J. Immunology, 184:5527-36.

Severson, K.M., Mallozzi, M., Driks, A., and Knight, K.L. (2010)  B cell development in GALT:  Role of bacterial and endogenous superantigen-like molecules, J. Immunology, 184:6782 – 6789.

Severson, K.M.,  Mallozzi, M., Bozue, J., Welkos, S, Cote, C.K., Knight, K.L., and Driks, (2009) A., Roles of the Bacillus anthracis Spore Protein ExsK in Exosporium Maturation and Germination. J. Bacteriology, 191:7587-7596.

Lanning, D.K., Severson, K.M., and Knight, K.L. (2008) Intestinal Bacteria: Mucosal Tissue Development and Gut Homeostasis, in Immunity against Mucosal Pathogens, Vajdy, M. ed., Springer Publ.  pp. 135-150.

Jasper,  P. J., Rhee, K. J., Kalis, S. L., Sethupathi, P., Yam P. C., Zhai, S. K., Knight, K. L.  B lymphocyte deficiency in IgH-transgenic rabbits. Eur J Immunol. 2007 Jul 12;37(8):2290-2299.

Kalis, S. L., Zhai, S.K., Yam, P. C, Witte, P. L., Knight, K. L. Suppression of B lymphopoiesis at a lymphoid progenitor stage in adult rabbits. Int. Immunol. 2007 Jun;19(6):801-11. 

Rhee, K.-J., Jasper, P.J., Sethupathi, P., Shanmugam, M., Lanning D. and Knight, K.L.: (2005) Positive selection of the peripheral B cell repertoire in gut associated lymphoid tissue.  J. Exptl. Med.201: 55-62.

Volgina, V., Yam, P. C., and Knight, K.L.: (2005) A negative regulatory elementin the rabbit 3'IgH chromosomal region. International Immunology, 17:973-982

Shanmugam, M., Sethupathi, P., Rhee, K.-J., Yong, S., and Knight, K.L. (2005) Bacterial-induced Inflammation in Germ-free Rabbit Appendix. Inflammatory Bowel Diseases. 11:992-996.

Lanning, D.K., Rhee, K.-J., and Knight, K.L.: (2005)  Intestinal Bacteria and Development of the B lymphocyte Repertoire, Trends in Immunology, 26:419

Mage, R.G., Lanning, D., and Knight, K.L.: (2005) B Cell and Antibody Repertoire Development in Rabbits: the Requirement of Gut-Associated Lymphoid Tissues, Development and Comparative Immunology. 30:137-153.

Jasper, P.J., Rhee, K.-J., Kalis, S.L., Sethupathi, P., Yam, P.-C., Zhai, S.-K., and Knight, K.L.: (2005) B Lymphocyte Deficiency in IgH Transgenic Rabbits,submitted.

Rhee, K.-J., Sethupathi, P., Driks, A., Lanning, D.K., Knight, K.L.: (2004) Role of Commensal Bacteria in Development of Gut-Associated Lymphoid Tissues and Preimmune Antibody Repertoire.J. Immunology, 172:1118.

Esteves, P.J., Lanning, D., Ferrand, N., Knight, K. L., Zhai, S.-K. and van der Loo, W.: (2004) Allelic variation at the VHa locus in natural populations of rabbit (Oryctolagus  cuniculus, L.), J. Immunology, 172:1044.

Lanning, D., Osborne, B.A. and Knight, K.L.: (2004) Immunoglobulin Genes and Generation of Antibody Repertoires in Higher Vertebrates: A Key Role for GALT, in Molecular Biology of B cells, edited by F.W. Alt, T. Honjo and M.S. Neuberger, Elsevier Science Ltd., 433-448.