F. Papola1, G. Fontecchio1, L. Ventura2, D. Adorno1, G. Fornaciari3

1Regional Centre of Immunohematology and Tissue Typing, ASL n.4, L’Aquila.
2Department of Pathology, San Salvatore Hospital, ASL n.4, L’Aquila.
3Department of Oncology, Transplants ad Advanced Technologies in Medicine, 
  Division of Paleopathology, University of Pisa, Via Roma 57, 56126 Pisa, Italy
  (e-mail: g.fornaciari@med.unipi.it)

Some paleopathological studies suggest that Rheumatoid Arthritis (RA) started in the New World, among ancient American Indians in Tennessee and neighbouring areas (5000-500 BC) and only after the discovery of America the disease was exported to the Old World.1

In 1996, in the Basilica of S. Francesco, famous for the frescoes of Piero della Francesca, a female mummy was discovered (figure, a). This 50-55 year old woman, nicknamed “Braids Lady”, lived in the 16th century and her body, well examined through several diagnostic instrumental investigations, shows many pathologic signs peculiar to RA: large erosions and evident subluxation of the metacarpophalangeal joints of the hands, lateral deviation of all the fingers with typical “z” deformation of the thumbs (figure, b), partial overlapping and fibular deviation of the toes, severe erosion of the right humerus head, but no involvement of the sacroiliac articulation, a prominent manifestation of  the ankylosing spondylitis.2

It is common knowledge that several autoimmune disorders are related to highly polymorphic and co-dominatly expressed antigens encoded by HLA (Human Leukocyte Antigen) complex. Particularly, the association between RA and some alleles of HLA-DRB1 locus, such as DRB1*0101, DRB1*0401, DRB1*0404, DRB1*0405, DRB1*0408, DRB1*1402 and DRB1*1001, is well established and a high percentage (> 90%) of individuals affected by this pathology possess some members of DRB1*01 or DRB1*04 gene families.3

The purpose of our work was to define the HLA-DRB genotype of “Braids Lady” for researching the possible presence of some RA susceptibility gene.

We first performed molecular  analysis on DNA obtained from deparaffinized tissue sections (by PCR-SSO) and then from the entire bone (by PCR-SSP).
HLA-DRB typing using PCR-SSO has shown the presence of DRB1*0101 and DRB1*1101 alleles (corresponding to DR1 and DR11 serotypes, respectively), together with positivity for DRB3 gene (phenotipically DR52). The assignment of allele DRB1*0101 has been confirmed by PCR-SSP (further details available from FP) (figure, c).


Figure legend

a: Natural mummy from the Basilica of S. Francesco in Arezzo (17th century). b: Left hand with large erosions of the metacarpophalangeal joints and lateral deviation of the fingers. c: HLA-DRB typing by PCR-SSP. The DNA, previously amplified using allele-specific primers, is run on 2% agarose gel stained with ethidium bromide and visualized on UV light. The bands (specific products), appearing in the gel at lanes 2 (200bp),  13 (270 bp) and 22 (240 bp), correspond to DRB1*0101, DRB1*11 and DRB3*01-03 alleles, respectively. The human growth hormone is added as internal control (1069 bp); the lane 1 is the negative control.
DRB1*0405 and *1001 are the most frequent in Asiatics with *1001 even in Spain;  DRB1*0101, together with DRB1*0401,*0404-05 sharing epitope Q(K/R)RAA, is associated with RA in Mediterranean patients, including Italians;4 finally, DRB1*1402, 0802, 0811 and 0407, absent or poorly represented in the Italian population, have generally a significantly higher gene frequency among Native Americans than other DRB1 alleles (Navajo and Pimans of Gila River Indian Community of Arizona, Lakota Sioux, Seri tribe of Northwest Mexico).5

In conclusion, we can state, by the positivity for DRB1*0101 allele in an Italian Renaissance mummy with clear “symptoms” of  Reumathoid Arthritis, that  this disease was present in the Old World before the discovery of America by Columbus in 1492.

1Rothschild BM, Woods RJ, Rothschild C, Sebes JI. Geographic distribution of rheumatoid arthritis in ancient North America: implication for pathogenesis. Semin Arthritis Rheum 1992; 22:181-7.
2Ciranni R, Garbini F, Neri E, Melai L, Giusti L, Fornaciari G. The “Braids Lady” of Arezzo: a case of rheumatoid arthritis in a 16th century mummy. Clin Exp Rheumatol 2002; 20: 745-52.
3Gonzalez-Gay MA, Garcia-Porrua C and Haajer AH. Influence of human leukocyte antigen-DRB1 on the susceptibility and severity of rheumatoid arthritis. Semin Atrhritis Rheum 2002;31:355-60.
4Bongi SM, Porfirio B, Rombola G, Palasciano A, Beneforti E, Bianucci G. Shared-epitope HLA-DRB1 alleles and sex ratio in Italian patients with rheumatoid arthritis. Joint Bone Spine. 2004;71:24-8.
5Leffell MS, Fallin MD, Hildebrand WH, Cavett JW, Iglehart BA, Zachary AA. HLA alleles and haplotypes among the Lakota Sioux: report of the ASHI minority workshops, part III. Hum Immunol 2004;65:78-89.