Within the framework of the Medici Project, a paleopathological team of experts from the University of Pisa, the University of Florence and the Superintendence for Florentine Museums, studied 20 tombs of some of the Medici family members (16th-18th centuries) housed in the socalled Medici Chapels of the Basilica of San Lorenzo in Florence. The project involves disciplines such as paleopathology, funerary archaeology, physical anthropology, palaeonutrition, parasitology, histology, histochemistry, immunohistochemistry, electron microscopy, molecular biology, and identification of ancient pathogens. The most recent biomedical technologies have been employed to obtain as much information as possible about the genetic make-up, eating habits, life styles and diseases of these important rulers of Renaissance Florence. Medical accounts and ancient autopsy reports imply that tertian malarial fevers caused the death of several members of the Medici family, such as Eleonora of Toledo (1522-1562), Cardinal Giovanni (1543-1562), don Garzia (1547-1562) and Grand Duke Francesco I (1531-1587). All members of the Medici family were hunted in the endemic malarial areas of Tuscany, such as the marshy areas surrounding their villas and along the swampy Maremma; therefore, they were highly exposed to the risk of being infected by falciparum malaria. To determine if malaria was the cause of death of those personages, we carried out immunological investigations and, then, compared the biological results to the historical sources. Bone samples were examined for the presence of Plasmodium falciparum histidine- rich- protein -2 (PfHRP2) and P. falciparum lactate dehydrogenase (PfLDH) using two different qualitative double–antibody immunoassays. Our findings provide the first modern laboratory evidence of the presence of P. falciparum ancient proteins in the skeletal remains of four members of the Medici family. We confirm the clinical diagnosis of the court physicians, using modern methods. Furthermore, immunological analyses detecting Leishmania infantum- specific peptides were carried out on the bone samples of the same four members of the Medici family. Positive identification by Western blot SDS-PAGE detected an IgG response anti- L. infantum in the human remains of Eleonora of Toledo (1522-1562), the wife of Cosimo I (1519-1574), suggesting that she had suffered from visceral leishmaniasis. This result was additionally supported by molecular ancient DNA (aDNA) analysis, able to identify a specific 120-bp fragment of a conserved region of the minicircle molecule of the parasite´s kinetoplastid mitochondrial DNA, which showed on direct sequencing a Leishmania specific sequence well compatible with L. Infantum. This is the first molecular and serological identification of L. infantum infection in a historic individual from southern Europe. Furthermore, we provide first evidence for visceral leishmaniasis (VL) and Mycobacterium tuberculosis complex (TB) co-infection in the skeletal remains of Eleonora of Toledo (1522 – 1562), wife of Cosimo I de’ Medici. Positive identification of Leishmania infantum in bone samples by Western blot SDS-PAGE detected an IgG response anti- L. infantum. PCR yielded successful amplification of a 121-bp fragment of a conserved region of the minicircle molecule of the parasite´s kinetoplastid mitochondrial DNA. Unique M. tuberculosis complex sequences (IS 6110 and rpoB) were also identified in Eleonora’s remains indicating that the historical reports which implied that, at age of 29, she developed pulmonary tuberculosis, were correct. Although distinct in aetiology and transmission mechanisms, VL and TB share several features. Many infections remain asymptomatic. Symptoms usually develop after several months or years in those who progress to clinical disease. The very long incubation periods may be related to immune suppression which apparently turns the latent infection into active disease. It has been suggested that TB represents one of the immunosuppressive conditions that can cause the progression of latent leishmanial infection to clinical leishmaniasis. Similarly, VL can reactivate a latent mycobacterial infection. Eleonora’s true state of health cannot now be evaluated. Long- term infections such as VL and TB pose many fundamental biological questions which are of great current interest in mammalian immune surveillance. The application of paleoimmunological techniques on ancient human bone tissue allowed the identification of ancient pathogens, such as malaria by Plasmodium falciparum and Leishmania infantum. Until now the immunocromatographic tests were applied only on mummified muscle. These results are of particular interest not only because they can confirm the historical data, as in the case of malaria, but also because they demonstrate that immunological tests can be applied to bone samples, opening new paths of research for the screening of large skeletal series. References FORNACIARI G., VITIELLO A., GIUSIANI S., GIUFFRA V., FORNACIARI A., VILLARI N., The Medici Project: first anthropological and paleopathological results of exploration of the Medici tombs in Florence. Med Secoli 2007; 19: 521-44. FORNACIARI G., GIUFFRA V., FERROGLIO E., BIANUCCI R., Malaria was the “killer” of Francesco I de’ Medici (1531-1587). Am J Med 2010; 123/6: 568-569. FORNACIARI G., GIUFFRA V., FERROGLIO E., GINO S., BIANUCCI R., Plasmodium falciparum immunodetection in the Medici Family of Florence (16th century). Trans R Soc Trop Med Hyg 2010; 104: 583-587.
Curtain raised on the Medici, Grand Dukes of Florence (16th - 17th centuries): New paleopathological results
FORNACIARI, GINO
2011-01-01
Abstract
Within the framework of the Medici Project, a paleopathological team of experts from the University of Pisa, the University of Florence and the Superintendence for Florentine Museums, studied 20 tombs of some of the Medici family members (16th-18th centuries) housed in the socalled Medici Chapels of the Basilica of San Lorenzo in Florence. The project involves disciplines such as paleopathology, funerary archaeology, physical anthropology, palaeonutrition, parasitology, histology, histochemistry, immunohistochemistry, electron microscopy, molecular biology, and identification of ancient pathogens. The most recent biomedical technologies have been employed to obtain as much information as possible about the genetic make-up, eating habits, life styles and diseases of these important rulers of Renaissance Florence. Medical accounts and ancient autopsy reports imply that tertian malarial fevers caused the death of several members of the Medici family, such as Eleonora of Toledo (1522-1562), Cardinal Giovanni (1543-1562), don Garzia (1547-1562) and Grand Duke Francesco I (1531-1587). All members of the Medici family were hunted in the endemic malarial areas of Tuscany, such as the marshy areas surrounding their villas and along the swampy Maremma; therefore, they were highly exposed to the risk of being infected by falciparum malaria. To determine if malaria was the cause of death of those personages, we carried out immunological investigations and, then, compared the biological results to the historical sources. Bone samples were examined for the presence of Plasmodium falciparum histidine- rich- protein -2 (PfHRP2) and P. falciparum lactate dehydrogenase (PfLDH) using two different qualitative double–antibody immunoassays. Our findings provide the first modern laboratory evidence of the presence of P. falciparum ancient proteins in the skeletal remains of four members of the Medici family. We confirm the clinical diagnosis of the court physicians, using modern methods. Furthermore, immunological analyses detecting Leishmania infantum- specific peptides were carried out on the bone samples of the same four members of the Medici family. Positive identification by Western blot SDS-PAGE detected an IgG response anti- L. infantum in the human remains of Eleonora of Toledo (1522-1562), the wife of Cosimo I (1519-1574), suggesting that she had suffered from visceral leishmaniasis. This result was additionally supported by molecular ancient DNA (aDNA) analysis, able to identify a specific 120-bp fragment of a conserved region of the minicircle molecule of the parasite´s kinetoplastid mitochondrial DNA, which showed on direct sequencing a Leishmania specific sequence well compatible with L. Infantum. This is the first molecular and serological identification of L. infantum infection in a historic individual from southern Europe. Furthermore, we provide first evidence for visceral leishmaniasis (VL) and Mycobacterium tuberculosis complex (TB) co-infection in the skeletal remains of Eleonora of Toledo (1522 – 1562), wife of Cosimo I de’ Medici. Positive identification of Leishmania infantum in bone samples by Western blot SDS-PAGE detected an IgG response anti- L. infantum. PCR yielded successful amplification of a 121-bp fragment of a conserved region of the minicircle molecule of the parasite´s kinetoplastid mitochondrial DNA. Unique M. tuberculosis complex sequences (IS 6110 and rpoB) were also identified in Eleonora’s remains indicating that the historical reports which implied that, at age of 29, she developed pulmonary tuberculosis, were correct. Although distinct in aetiology and transmission mechanisms, VL and TB share several features. Many infections remain asymptomatic. Symptoms usually develop after several months or years in those who progress to clinical disease. The very long incubation periods may be related to immune suppression which apparently turns the latent infection into active disease. It has been suggested that TB represents one of the immunosuppressive conditions that can cause the progression of latent leishmanial infection to clinical leishmaniasis. Similarly, VL can reactivate a latent mycobacterial infection. Eleonora’s true state of health cannot now be evaluated. Long- term infections such as VL and TB pose many fundamental biological questions which are of great current interest in mammalian immune surveillance. The application of paleoimmunological techniques on ancient human bone tissue allowed the identification of ancient pathogens, such as malaria by Plasmodium falciparum and Leishmania infantum. Until now the immunocromatographic tests were applied only on mummified muscle. These results are of particular interest not only because they can confirm the historical data, as in the case of malaria, but also because they demonstrate that immunological tests can be applied to bone samples, opening new paths of research for the screening of large skeletal series. References FORNACIARI G., VITIELLO A., GIUSIANI S., GIUFFRA V., FORNACIARI A., VILLARI N., The Medici Project: first anthropological and paleopathological results of exploration of the Medici tombs in Florence. Med Secoli 2007; 19: 521-44. FORNACIARI G., GIUFFRA V., FERROGLIO E., BIANUCCI R., Malaria was the “killer” of Francesco I de’ Medici (1531-1587). Am J Med 2010; 123/6: 568-569. FORNACIARI G., GIUFFRA V., FERROGLIO E., GINO S., BIANUCCI R., Plasmodium falciparum immunodetection in the Medici Family of Florence (16th century). Trans R Soc Trop Med Hyg 2010; 104: 583-587.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
