Cross section through bone marrow - negative control

A cross section through a femur from a control, non-transgenic mouse. Compare this background fluorescence to the green fluorescence expressed by B lineage progenitors expressing rag2-GFP in slide 4.

B cell development in situ

A cross section through a murine femur reveals B lineage progenitors nestled within the bone cavity.

Lymphocytes within the bone marrow (upper right) are outlined in faint green; greenish line at lower left is edge of bone.

Tracing rag2-GFP expression in T cells

Green fluorescence marks T cell progenitors undergoing V(D)J recombination to assemble a T cell receptor. Slide 6 shows this same section under transmitted light. Note that virtually all T cells express rag2.

Compare to rag2 expression in bone marrow B cells, slide 4.

Tracing rag2-GFP expression in B cells

Dim green fluorescence marks B lineage progenitors (middle and upper right) undergoing V(D)J recombination to make antibodies. Slide 2 show this B cell section under transmitted light, and slide 1 shows a comparable image from a control mouse that does not express rag2-GFP.

Note that rag2+ B lymphocytes in bone marrow are rare as compared to rag2+ T cells in the thymus (slide 3). This is because B cell precursors are not the only hematopoietic subset in the bone marrow while T cell precursors form the bulk of thymus cells.

developing B lymphocytes

B lineage progenitors (small, round) receive critical developmental signals from bone marrow stromal cells (larger, adherent).

thymus cross section

The thymus is packed full of developing T lineage progenitors. Visualize which cells are in the process of making a T cell receptor by seeing this same section with fluorescent imaging in slide 3.

Lisa Borghesi, PhD

Lisa is a B cell biologist with research interests in hematopoiesis, V(D)J rearrangement, and cancer. Her work spans basic aspects of lymphocyte development in the mouse model and translational work on cancers of B lineage origin in man. She combines expertise in molecular and cellular biology to characterize the transcription factors that guide the specification of uncommitted hematopoietic progenitors to the B lymphocyte lineage and how this transcriptional cascade is perturbed in disease states.

Qi Yang, MD, fourth year graduate student

Qi comes to us from Tongji Medical College, China. Qi is a skilled molecular biologist who is studying the mechanisms that regulate the functional integrity of stem cells. Her first-authored paper was published in the November 2008 issue of JI, and featured prominently in oral presentations at both Keystone and AAI meetings in 2009. And, oh yes, she's about to have a baby - our first lab baby!!!

Patricia Santos, BS, third year graduate student

Patti, a native of the Philippines (yes, Pgh is on the chilly side), is interested in the control of cell cycle regulation. Excessive cell cycling or loss of stem cell quiescence is linked to impaired self-renewal of HSCs, malignant transformation, and even hematopoietic failure. Patti is studying the role of the transcription factor E47 in the regulation of HSC proliferation, and the specific target genes through which E47 acts in vivo.

Lela Kardava, PhD, postdoctoral fellow 2006-2008, now doing second postdoc at the NIH

Lela has a long-standing interest in the B cell cancer chronic lymphocytic leukemia (CLL), in part because of the high incidence of CLL in her home country of Georgia. Here, Lela's work focused on the transcription factor E2A/E47 as an essential target gene in survival and proliferation of primary human CLL cells. Currently at the NIH under Tony Fauci's umbrella, Lela focuses on B cell activities in HIV patients.

Kristy Pilbeam, research associate 2005-2007, now in medical school class of 2011

Kristy's interest in cancer biology led her to examine the biological consequences of V(D)J rearrangements in NK cell precursors to mature NK cell activity in normal and disease circumstances. Her first-authored paper was published in JI. Kristy is now doing quite well in her studies at Lake Erie College of Medicine and is tackling her first clinical rotations. She looks forward (?) to Step II of the boards.

Jen Aites, undergrad researcher then tech 2005-2006, currently in medical school class of 2010

Jen's work on V(D)J recombination in B cells earned her co-authorship on our J Exp Med paper. Now in her last year of med school at Penn State, Jen is preparing for residency. Second most recent update from her: "I'm rather disappointed with the lack of cool immunologic disorders and the overwhelming amount of runny noses and coughs." Most recent update: "I had my meeting this week with one of the deans to go through all of my records...It's amazing the things the school knows about me."

Tony St Leger, undergraduate researcher 2007-2008, now second year graduate student at Pitt

Tony's experience as a two time Science Olympic participant and plant pollination biologist - humorously self-described as a glorified bee - says it all. While in the lab, Tony got a student scholarship for his senior year and co-authorship on 2008 JI paper. Tony has officially joined Bob Hendrick's lab for his thesis work, an experience he finds far more predictable than the nuances of home ownership. Congrats to Tony and Kerry on their recent nuptials!

Mike Magee, student researcher 2005-2006, now second year graduate student at Thomas Jefferson U

Mike worked with us throughout his junior and senior years at Pitt, defining the expression of the transcription factor E47 in B cells from normal and diseased patients. Following a year of postgraduate research, Mike jumped into grad school where his thesis project in Scott Waldman's lab focuses on adenoviral based cancer vaccine targeting.

Yanille Scott, summer student 2006, now first year graduate student at Pitt

Yanille's summer project focused on the molecular regulation of transcription factors that are important for B cell development. After graduating from Benedict College and then working in pharma for a bit, we have successfully lured her back to Pitt. Yanille has just started her first year in the PhD program in Infectious Disease and Microbiology.

Shakira Nelson, summer student 2005, currently second year graduate student at Penn State

In her summer project, Shakira showed that the transcription factor E47 is required to initiate V(D)J recombination in B cells. Following a stint on flu vaccine work with the US Dept of Health & Human Services, Shakira is excelling in the PhD program in Pathobiology at Penn State. Most recent update: "Campus life is not as weird as I thought it would be."

Jason Heakins, undergraduate researcher spring 2007

Using his experience in the chemistry department as a springboard, Jason applied his research skills to a biological problem. His work focused on characterizing a specific gene pathway that regulates hematopoietic stem cell function and longevity. Having several AP credits under his belt positioned Jason well for early graduation.

Elizabeth Crowley, undergraduate researcher fall 2006

While in the lab, Elizabeth studied one molecular mechanism underlying chromosomal translocation. Her worked led to key figures in Kristy's 2008 JI paper. Elizabeth was working with the Allegheny County Health Department last we knew.

Preslav Lefterov, lab set up fall 2004

Preslav, known to many as the mysterious Bulgarian, joined the lab before there even was a lab. While helping to build the lab and seed lab philosophy, Preslav performed construction on his dream - establishing Machine Age recording studio.